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Technicians

Collection Instructions

Collection and Transport of Specimens

A. Tissue Preparation for General Histology

The most important step in the histological exam is Fixation. If a specimen is improperly fixed, the histology of the tissue is compromised or lost. Because many histologic samples cannot be recovered or repeated, it is critical for everyone involved with the tissue specimen to have a working understanding of fixation. Expected routine TAT is 24-48 hours.

Fixatives of choice for Histology are:

  • 10% Neutral Buffered Formalin for transportation from Collecting facilty to Laboratory.
  • Bouin's fixative for Bone Marrow specimen processing.
  • RPMI for specimens (e.g. Lymph Nodes) processed for genetic testing
  • Saline for fresh specimens
  • CytoRich Red for fluid specimens

1. Fixatives are used to:

  • a. Prevent autolysis (cell self destruction from enzymes)
  • b. Prevent putrefaction (bacterial destruction of the cell)
  • c. To harden the tissue (make tissue easier to handle)
  • d. To kill infectious agents.

2. In order to maximize accurate results for tissue examination, the person handling the original specimen should:

  • a. Pre-label the specimen containers with Patient Name, Collection Date, and Collection Site.
  • b. Put the specimen in fixative as soon as it is removed from the patient. This must not wait until the procedure is over. Small specimens are especially susceptible to autolysis and putrefaction.
  • c. Completely cover the specimen with formalin. A 1:20 ratio (size of specimen to formalin) is ideal. This is especially crucial for large specimens.
  • d. Seal the container and check for leaks. Formalin is a caustic chemical and should be handled with care. Double check containers to confirm they are all labeled with the Patient Name, Collection Date, and Collection Site.
  • e. Fill out the Pathology Requisition.
  • f. Place the container with the requisition in a specimen transportation bag and place in the designated specimen pick-up area.
  • g. According to the facility’s courier set-up, call for a pick-up or wait for the scheduled pick-up time.

3. For Amputation of Large Appendages

  • Cover the transected bone with an absorbent pad (e.g. thick gauze, cloth), to prevent sharp edges from perforating the containing bag.
  • Place the appendage inside a red biohazard bag (or similar transport bag), and wrap tightly around the appendage. Tape firmly closed.
  • Label the wrapped appendage with patient information. If this stage is not labeled, the specimen is not considered properly labeled by CAP and CLIA regulations.
  • Place the wrapped appendage into a brown biohazard box. Tape firmly closed.
  • Place the box inside another red biohazard bag. Tie or tape closed.
  • Make sure the completed requisition and accompanying paperwork are with the specimen. It is preferred to put the paperwork in a small bag and tape that bag to the bagged box containing the appendage.
  • Have the specimen in the designated pick-up area for the courier to retrieve.

If refrigeration is available, refrigerate the appendage at 4°C until courier pick-up.

Note: If the patient or family would like to request this appendage to be returned for burial, make a note on the requisition stating the request and contact information for the hospital staff member to contact. If a note is not made on the requisition, laboratory staff may not store the appendage in a manner appropriate for burial.

See sections U-W for more information.

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B. Fine Needle Aspiration (FNA)

Expected Routine TAT 24-48 hours

1. The FNA Kit consists of:

  • Three 50mL tubes, partially-filled with CytoRich Red fluid
  • Each tube is for a single specimen, allowing for three cases per kit.

2. Fixatives are used to:

  • a. Prevent autolysis (cell self destruction from enzymes)
  • b. Prevent putrefaction (bacterial destruction of the cell)
  • c. To harden the tissue (make tissue easier to handle)
  • d. To kill infectious agents.
  • e. Prevents the tissue from drying out.

3. FNA Collection:

  • a. Estimate the amount of the Specimen and pour out any excess CytoRich Red from the 50mL tube.
    tube.
  • b. Eject the Specimen into the 50mL container of CytoRich Red, maintaining a 50/50 ratio of specimen to fixative.
  • c. A minimum of 10mL of Specimen is preferred. Sending the Specimen in full is ideal.

* Note: This method can be used for an FNA of any site. If a core biopsy is collected, insert it into a 20mL formalin vial and follow normal biopsy collection procedures. *Do not send sharps to the laboratory*.

4. In order to maximize accurate results for cytology testing, the person collecting and handling the original specimen should:

  • a. Pre-label all FNA Kit containers and slides.
  • b. Seal the container(s) after specimen has been inserted and check for leaks. Double check the containers and slides to confirm they are all labeled with the Patient Name, Collection Date, and Collection Site.
  • c. Fill out the Pathology Requisition.
  • d. Place the container(s) in the sealable portion of a specimen transportation bag and place in the requisition in the side pocket. Then place specimen transport bag in designated specimen pick-up area.
  • e. According to the facility’s courier set-up, call for a pick-up or wait for the scheduled pick-up time.


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C. Urine Specimen Preparation for Cytology

Expected Routine TAT 24-48 hours

1. The fixative of choice is reagent Alcohol.

2. Fixatives are used to:

  • a. Prevent autolysis (cell self destruction from enzymes)
  • b. Prevent putrefaction (bacterial destruction of the cell)
  • c. To harden the tissue (make tissue easier to handle)
  • d. To kill infectious agents.
  • e. Prevents the tissue from drying out.

3. Urine Specimen Preparation for Cytologic Review

  • a. Have the patient urinate into a urine catch cup labeled with their name.
  • b. Receive the specimen from the patient in the labeled catch cup.
  • c. Pre-label the specimen transportation container with the Patient Name, Collection Date, and
    “Urine” for the Site.
  • d. Pour the specimen into the Pre-labeled specimen transportation container, leaving room for the
    fixative to be added. Specimen/Fixative ratio should be 1:1, (equal amounts of each).
  • e. Pour fixative (50% Alcohol) into specimen transportation container, make sure ratio of
    specimen to fixative is 1:1.
  • f. Apply the lid on the transport container and tighten. Double check specimen transport
    container for Patient Name, Collection Date, and “Urine” as Collection Site.
  • g. Completely fill out the Precision Pathology Requisition.
  • h. Put specimen and requisition in specimen bag for transportation.
  • i. According to the facility’s courier set-up, call for a pick-up or wait for the scheduled pick-up
    time.

*NOTE*: Any questions regarding a specimen or its fixation should be directed to the Laboratory Director.

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D. All Other Body Fluids

Expected Routine TAT 24-48 hours

  • The fixative of choice is Cyto Rich Red.
    Note: Fresh specimens for HIV, TB, HepB or HepC are not accepted at the Laboratory. The specimen must be placed in fixative.
  • Fixative should be added to all fluid immediately after collection for optimal results.
    1. Estimate the amount of the Specimen and pour out any excess CytoRich Red from the 50mL tube to create a 50/50 ratio.
    2. Eject the Specimen into the 50mL container of CytoRich Red, maintaining a 50/50 ratio of specimen to fixative.
    3. A minimum of 30mL of Specimen is preferred. Sending the entire Specimen is ideal.
  • For specimens sent to the Laboratory the same day, and/or very large amounts of specimen (e.g. 1L vacutainers), the specimen may be sent fresh. The specimen is to be kept refrigerated until pick-up.
  • In order to maximize accurate results for cytology testing, the person collecting and handling the original specimen should:
    1. a. Pre-label all FNA Kit containers and slides. Individual slides must be labeled with a minimum of one patient identifier, and the slide holder must be labeled with a minimum of two patient identifiers. (e.g. Full Name and Birthdate)
    2. b. Seal the container(s) after specimen has been inserted and check for leaks. Double check the containers and slides to confirm they are all labeled with the Patient Name, DOB, Collection Date, and Collection Site.
    3. c. Fill out the Pathology Requisition.
    4. d. Place the container(s) in the sealable portion of a specimen transportation bag and place in the requisition in the side pocket. Then place specimen transport bag in designated specimen pick-up area.
    5. e. According to the facility’s courier set-up, call for a pick-up or wait for the scheduled pick-up time

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E. GYN Cytology Collection

1. Amplified Chlamydia / Gonorrhea – Swab Specimens

Test Code(s): [SWAB CT], [SWAB CT/GC COMBO]

a. Purpose of Test

The BDProbeTec™ ET system is designed to detect the presence of Chlamydia trachomatis and Neisseria gonorrhoeae in endocervical swabs, male urethral swabs and male and female urine specimens using the appropriate collection method. The CULTURETTE™ DIRECT Cleaning – Collection and Transport System and the BD Mini-Tip CULTURETTE™ DIRECT Collection and Transport System are the devices that can be used to collect swab specimens for testing on the BDProbeTec™ ET Instrument. Urine specimens must be collected in a BD Vacutainer™, plastic preservative-free, specimen collection cup.

b. Collection Device/Specimen Type0

  • i. CDD Supplied Wet Transport Collection Kit (Preferred)
  • ii. BD ProbeTec ET female endocervical dry swab collection kit
  • iii. BD ProbeTec ET male urethral/conjunctival mini swab collection kit.

c. Collection Procedure (Female Endocervical Swab)

  • i. Remove excess mucus from the cervical os with the large (orange shaft) cotton-tipped cleaning swab provided in the Swab Transport Kit and discard.
  • ii. Insert the polyester Female Collection Swab (white shaft) into the cervical canal and rotate for 15-30 seconds.
  • iii. Withdraw the swab carefully; avoid contact with the vaginal mucosa.
  • iv. Immediately place the swab into the transport tube.
  • v. Break the shaft at the score line.

NOTE: Do not submit specimens using the large (orange shaft) swab. Specimens will be rejected.

  • vi. Screw the cap firmly onto the specimen tube.
  • vii. Completely fill out the Precision Pathology OBGYN Requisition.
  • viii. Put the specimen and requisition in a specimen bag for transportation.
  • ix. According to the facility’s courier set-up, call for a pick-up or wait for the scheduled pick-up time

d. Collection Procedure (Male Urethral Swab)

  • i. Use the BD Mini-Tip CULTURETTE™ DIRECT Collection and Transport System – Male (Blue Cap).
  • ii. Insert the male swab 2-4cm into the urethra and rotate for 3-5 seconds.
  • iii. Withdraw the swab and place the swab into the transport tube.
  • iv. Completely fill out the Precision Pathology OBGYN Requisition.
  • v. Put the specimen and requisition in a specimen bag for transportation.
  • vi. According to the facility’s courier set-up, call for a pick-up or wait for the scheduled pick-up time

e. Specimen Storage and Transport

  • i. When stored at 2-27°C, the CULTURETTE™ DIRECT collection swab and the Mini-Tip CULTURETTE™ DIRECT swabs are stable for 4-6 days or 30 days when stored at 2-8°C.
  • ii. Ship to CDD at room temperature.
  • iii. Place the specimen in a CDD transport container.
  • iv. Secure transport container with a rubber band.
  • v. Place the container in an opaque ziplock bag.
  • vi. Place the closed Ziplock bag in a Lab Pack (white).
  • vii. Remove the red adhesive strip from the Lab Pack.
  • viii. Seal securely and tightly.
  • ix. Completely fill out the Precision Pathology OBGYN Requisition.
  • x. Put the specimen and requisition in a specimen bag for transportation.
  • xi. According to the facility’s courier set-up, call for a pick-up or wait for the scheduled pick-up time

f. Procedural Notes

All results, negative and positive, are reported the same day that the specimen is received in the laboratory (Monday through Friday).

g. CPT Codes

  • i. CT 87491
  • ii. GC 87591

h. Reference Values

  • i. Chlamydia Negative
  • ii. Gonorrhea Negative

2. Amplified Chlamydia/Gonorrhea – Urine Specimens

Test Code(s): [URINE CT], [URINE CT/GC COMBO]

a. Purpose of Test

The BDProbeTec™ ET system is designed to detect the presence of Chlamydia trachomatis and Neisseria gonorrhoeae in endocervical swabs, male urethral swabs and male and female urine specimens using the appropriate collection method. The CULTURETTE™ DIRECT Cleaning – Collection and Transport System and the BD Mini-Tip CULTURETTE™ DIRECT Collection and Transport System are the devices that can be used to collect swab specimens for testing on the BDProbeTec™ ET Instrument. Urine specimens must be collected in a BD Vacutainer™, plastic preservative-free, specimen collection cup.

b. Collection Device / Specimen Type

  • i. BD Vacutainer™ Urine Collection Kit
  • - Sterile, plastic, preservative-free specimen collection cup.
  • - Plain UA Tube (Yellow Top)
  • ii. Unpreserved Urine in Sterile (Yellow Top) tube

c. Collection Procedure (Male / Female Urine)

  • i. The patient should not have urinated for at least one hour prior to specimen collection.
  • ii. Collect the specimen in a sterile, plastic, preservative-free specimen collection cup and replace the lid. (BD Vacutainer™)
  • iii. The patient should collect the first 15-60mL of voided urine (the first part of the stream- NOT midstream)
  • iv. Clinic Instruction: Peel back, but do not remove, the protective sticker to expose the rubber-covered cannula.
  • v. Push the plain UA Tube (yellow top) into the integrated transfer port. Hold in position until the flow of urine stops. Remove the tube.
  • vi. Place the protective sticker back over the integrated transfer port.
  • vii. Completely fill out the Precision Pathology OBGYN Requisition.
  • viii. Put the specimen and requisition in a specimen bag for transportation.
  • ix. According to the facility’s courier set-up, call for a pick-up or wait for the scheduled pick-up time

d. Urine Specimen Storage and Transport

  • i. Specimens may be stored at 2-8°C for up to seven days or at 2-30°C for up to 30 hours before testing.
  • ii. Ship to CDD at room temperature.
  • iii. Place the specimen in a CDD transport container.
  • iv. Secure transport container with a rubber band.
  • v. Place the container in an opaque ziplock bag.
  • vi. Place the closed Ziplock bag in a Lab Pack.
  • vii. Remove the red adhesive strip from the Lab Pack
  • viii. Seal securely and tightly.
  • ix. Completely fill out the Precision Pathology OBGYN Requisition.
  • x. Put the specimen and requisition in a specimen bag for transportation.
  • xi. According to the facility’s courier set-up, call for a pick-up or wait for the scheduled pick-up time

e. Procedural Notes

All results, negative and positive, are reported the same that the specimen is received in the laboratory (Monday through Friday).

f. CPT Codes

  • i. CT 87491
  • ii. GC 87591

g. Reference Values

  • i. Chlamydia Negative
  • ii. Gonorrhea Negative

3. Syphilis Testing RPR screen, FTA-ABS, or CAPTIA Syphilis-IgG

Test Code(s): [SYPH], [SYPH-HIV]

a. Purpose of the Test

  • i. Syphilis tests detect antibodies to the bacterium that causes syphilis (Treponema pallidum) in blood, body fluid or tissue. The tests are used to screen for or to confirm a syphilis infection.
  • ii. Rapid plasma reagin (RPR) test: The RPR test also detects syphilis antibodies.
  • iii. Enzyme immunoassay (EIA) test: This is a newer blood test that checks for antibodies to the bacteria that cause syphilis. A positive EIA test should be confirmed with either the VDRL or RPR tests.
  • iv. Fluorescent treponemal antibody absorption (FTA-ABS) test: The FTA-ABS test detects antibodies to the bacteria that cause syphilis and can be used to detect syphilis except during the first three to four weeks after exposure to syphilis bacteria. It is more difficult to do and may be used to confirm a syphilis infection after another method tests positive for the syphilis bacteria.

b. Collection Device / Specimen Type

  • i. Plastic Red Top (no additive) blood collection tube.
  • ii. Minimum of 1mL serum

c. Collection Procedure

  • i. Following the standard phlebotomy venipuncture technique, blood is drawn from a vein, usually from the inside of the elbow.
  • ii. Collect at least 4mL of whole blood in a plastic Red Top blood collection tube.
  • iii. Label the specimen container with patient’s name and at least one other identifier (e.g. birth date)
  • iv. Completely fill out the Precision Pathology OBGYN Requisition.
  • v. Put the specimen and requisition in a specimen bag for transportation.
  • vi. According to the facility’s courier set-up, call for a pick-up or wait for the scheduled pick-up time

d. Procedural Notes

  • i. All specimens that are RPR negative shall be retained for at least fourteen days after the test result is reported.
  • ii. All specimens tested as RPR or FTA positive shall be retained for at least 180 days after the test result is reported.
  • iii. Specimens initially reactive by Captia IgG EIA method are retested by RPR.
  • iv. Samples that are reactive by both screening methods are confirmed by FTA-ABS.
  • v. Initial screen and RPR, if required, are performed the same day the sample is received at CDD.
  • vi. Hemolysis of blood specimens adversely affects results. Hemolyzed specimens will be rejected for testing.

e. CPT Codes

  • i. RPR 86592
  • ii. RPR Titer 86593
  • iii. FTA-ABS 86781
  • iv. SYPHILIS EIA 86318

f. Reference Values

SYPHILIS Negative

4. HIV-1/2 Plus O Blood Specimens

Test Code(s): [EIA HIV], [SYPH-HIV]

a. Purpose of Test

The test is used for the qualitative detection of antibodies to HIV-1/2 Plus O virus in human serum and plasma samples. Specimens initially reactive will be re-tested in duplicate. Those specimens testing positive on a subsequent test (repeatedly reactive) will be tested using a more specific Western Blot test.

b. Collection Device / Specimen Type

  • i. Plastic Red Top (no additive) blood collection tube.
  • ii. Minimum 1mL serum.

c. Collection Procedure

  • i. Following the standard phlebotomy venipuncture technique, blood is drawn from a vein, usually from the inside of the elbow.
  • ii. Collect at least 4mL of whole blood in a plastic Red Top blood collection tube.
  • iii. Label the container with the patient’s name and at least one other identifier (e.g. birth date)
  • iv. Completely fill out the Precision Pathology OBGYN Requisition.
  • v. Put the specimen and requisition in a specimen bag for transportation.
  • vi. According to the facility’s courier set-up, call for a pick-up or wait for the scheduled pick-up time

d. Procedural Notes

  • i. For long-term storage, specimens must be frozen at = -20°C.
  • ii. Initial screen and repeat, if required, are performed the same day the sample is received in the laboratory.
  • iii. Western Blots are performed within three working days of receiving the intial sample.

e. CPT Codes

HIV-1/2 Plus O Negative

f. Reference Values

HIV-1/2 Plus O Negative

5. Conventional Pap Smear

Test Code(s): [PAP]

a. Purpose of Test

The Pap Smear is a screening technique to assist in the detection of premalignant and malignant lesions of the cervix.

b. Collection Device / Specimen Type

  • i. Use the “No Touch®” One-Slide Pap Smear Kit.
  • ii. The kit contains all materials needed to provie a conventional Pap smear (wooden spatula, cotton-tipped swab and glass slide).

c. Collection Procedure

  • i. Do not remove the slides from the kit packet. Sections “V”, “C” and “E” are noted on the slide when the slide is enclosed in the kit packet.
  • ii. Neatly print the patient’s name and identifying number in pencil on the frosted end of the slide.
  • iii. Obtain a vaginal smear with the flat (non-notched) end of the wooden spatula. Obtain a hormonal evaluation sample by lightly scraping the mid-lateral vaginal wall. Spread material evenly on section “V” of the slide and immediately wet the smear area with fixative, covering sections “C” and “E”.
  • iv. Obtain a cervical smear by rotating the notched end of the wooden spatula around the ectocervix. Spread the material evenly on section “C” and immediately wet the smear area with fixative, covering section “E” as shown in the kit packet.
  • v. Obtain an endocervical smear by gently inserting the endocervical brush, or cotton-tipped swab, into the endocervical canal and rotating at least one full turn (360°). Gently remove the endocervical brush, or swab, without touching any vaginal surfaces. Spread the material evenly by rolling the endocervical brush, or swab, back and forth over section “E” and wet immediately with fixative.
  • vi. Close the No Touch® slide packet and label with patient’s name and at least one other identifier (e.g. birth date).
  • vii. Completely fill out the Precision Pathology OBGYN Requisition.
  • viii. Put the specimen and requisition in a specimen bag for transportation.
  • ix. According to the facility’s courier set-up, call for a pick-up or wait for the scheduled pick-up time

d. Procedural Notes

  • i. Do not use the endocervical brush on pregnant patients to sample the endometrium, or on inflamed or infected tissues.
  • ii. The PAP smear is not a diagnostic test. It is only a screening procedure designed to detect cancerous and pre-cancerous lesions.
  • iii. The name and identifying number must be written on the slide in pencil in order for the slide to be identify in case it is separated from the slide mailer.
  • iv. The information written on the slide must match the requisition. Specimens with discrepancies between the slide and requisition will be rejected for testing and returned to the submitting clinic for final disposition.

e. CPT Code

88164

f. Reference Values

Interpretive Report

6. SurePath® Pap Test

a. Purpose of Test

The purpose of the BD SurePath™ liquid-based Pap test is a screening technique to assist in the detection of premalignant and malignant lesions of the cervix.

b. Collection Device / Specimen Type

  • i. SurePath™ Pap Test Vial
  • ii. Use either a broom-like device with the detachable head, or the SurePath™ brush/spatula collection device.

c. Collection Procedure – Broom-like Device

  • i. Obtain an adequate sampling from the cervix using a broom-like device with detachable head. Insert the central bristles of the broom into the endocervical canal deep enough to allow the shorter bristles to fully contact the ectocervix. Push gently, and rotate the broom in a clockwise direction five times.
  • ii. Using the thumb and forefinger of gloved hand(s), disconnect the head of the device from the handle, and insert the head into the collection vial. Discard the handle of the sampling device. Do not touch the head of the device.
  • iii. Specimens received without the head of the collection device will be rejected.
  • iv. Tight the cap securely. Specimens that leak in transit cannot be tested.
  • v. Label the vial with the patient’s name and at least one other patient identifier (e.g. birth date)
  • vi. Completely fill out the Precision Pathology OBGYN Requisition.
  • vii. Put the specimen and requisition in a specimen bag for transportation.
  • viii. According to the facility’s courier set-up, call for a pick-up or wait for the scheduled pick-up time

d. Collection Procedure - SurePath™ Brush and Spatula

  • i. Obtain an adequate sampling from the ectocervix using a plastic spatula with detachable head.
  • ii. Using the thumb and forefinger of a gloved hand, disconnect the head of the device from the handle by breaking at the score line. Insert the head into the collection vial. Discard the handle of the sampling device. Do not touch the head of the device.
  • iii. Obtain an adequate sampling from the endocervix using an edocervical brush device with detachable head. Insert the brush into the cervix until only the bottommost fibers are exposed. Slowly rotate ¼ or ½ turn in one direction. DO NOT OVER-ROTATE.
    NOTE: More is not better. Over-rotating the endocervical brush is a major source of error when collecting the SurePath™ liquid-based Pap test. In an effort to ensure adequate sampling of the endocervix, many clinicians over-rotate the brush, which results in the endocervical cells coming off the brush. This often results in “No endocervical/transition zone component identified.”
  • iv. Using the thumb and forefinger of a gloved hand, disconnect the head of the device from the handle by breaking at the score line. Insert the head into the collection vial. Discard the handle of the sampling device. Do not touch the head of the device.
  • v. Specimens received without the head of the collection device will be rejected.
  • vi. Tighten the cap securely. Specimens that leak in transit cannot be tested.
  • vii. Label the vial with the patient’s name and at least one other patient identifier (e.g. birth date)
  • viii. Completely fill out the Precision Pathology OBGYN Requisition.
  • ix. Put the specimen and requisition in a specimen bag for transportation.
  • x. According to the facility’s courier set-up, call for a pick-up or wait for the scheduled pick-up time

e. Procedural Notes

  • i. Do not use the Endocervical brush on pregnant patients, to sample the endometrium, or on inflamed or infected tissues.
  • ii. SurePath™ is not a diagnostic test. It is only a screening procedure designed to detect cancerous and pre-cancerous lesions.

f. CPT Code(s)

88142

g. Reference Values

Interpretive Report

7. ThinPrep® Pap Test

Test Code(s): [THINPREP], [TP RFX HPV], [TP-HPV]

a. Purpose of Test

The Cytyc ThinPrep™ liquid-based Pap test is a screening technique to assist in the detection of premalignant and malignant lesions of the cervix.

b. Collection Device/Specimen Type

  • i. ThinPrep™ Pap Test Vial
  • ii. Use either a broom-like device, or a brush/spatula collection device.

c. Collection Procedure – Broom-like Device

  • i. Obtain an adequate sampling from the cervix using a broom-like device. Insert the central bristles of the broom into the endocervical canal deep enough to allow the shorter bristles to fully contact the ectocervix. Push gently, and rotate the broom in a clockwise direction five times.
  • ii. Rinse the broom as quickly as possible into the PreservCyt® solution vial by pushing the broom into the bottom of the vial at least ten times, forcing the bristles apart. As a final step, swirl the broom vigorously to further release material. Discard the collection device.
  • iii. Tighten the cap so that the torque line on the cap passes the torque line on the vial.
  • iv. Label the vial with the patient’s name and at least one other patient identifier (e.g. birth date)
  • v. Completely fill out the Precision Pathology OBGYN Requisition.
  • vi. Put the specimen and requisition in a specimen bag for transportation.
  • vii. According to the facility’s courier set-up, call for a pick-up or wait for the scheduled pick-up time

d. Collection Procedure – ThinPrep™ Brush and Spatula

  • i. Obtain an adequate sampling from the ectocervix using a plastic spatula.
  • ii. Rinse the spatula as quickly as possible into the PreservCyt® solution vial by swirling the spatula vigorously in the vial at least ten times. Discard the spatula.
  • iii. Obtain an adequate sampling from the endocervix using an endocervical brush device. Insert the brush into the cervix until only the bottommost fibers are exposed. Slowly rotate ¼ or ½ turn in one direction. DO NOT OVER-ROTATE.
  • NOTE: More is not better. Over-rotating the endocervical brush is a major source of error when collecting the ThinPrep™ liquid-based Pap test. In an effort to ensure adequate sampling of the endocervix, many clinicians over-rotate the brush, which results in the endocervical cells coming off the brush. This often results in “No endocervical/transition zone component identified.”
  • iv. Rinse the brush as quickly as possible in the PreservCyt® solution by rotating the device in the solution at least ten times while pushing against the PreservCyt® vial wall. Swirl the brush vigorously to further release material. Discard the brush.
  • v. Tighten the cap so that the torque line (black mark) on the cap passed the torque line on the vial.
  • vi. Label the vial with the patient’s name and at least one other patient identifier (e.g. birth date)
  • vii. Completely fill out the Precision Pathology OBGYN Requisition.
  • viii. Put the specimen and requisition in a specimen bag for transportation.
  • ix. According to the facility’s courier set-up, call for a pick-up or wait for the scheduled pick-up time

e. Procedural Notes

  • i. Do not use Endocervical brush on pregnant patients, to sample the endometrium, or on inflamed or infected tissues.
  • ii. ThinPrep™ is not a diagnostic test. It is only a screening procedure designed to detect cancerous and pre-cancerous lesions.

f. CPT Code(s)

88142

g. Reference Values

Interpretive Report

8. HPV, Cervista™ HPV HR

Test Code(s): [TPREP HPV], [SPATH HPV], [DIGENE HPV]

a. Purpose of Test

Approximately forty of the over 100 documented HPV types infect the anogenital area and are transmitted sexually. Anogenital HPV is associated with virtually all cancers of the cervix. Cervical cancer has previously been shown to be highly preventable when cytological and HPV screening programs are employed to facilitate the detection and treatment of pre-cancerous lesions.

b. Collection Device / Specimen Type

  • i. Residual PreserveCyt fluid for ThinPrep® samples
  • ii. Residual SurePath® fluid for SurePath® samples
  • iii. Digene® cervical sampler

c. Collection Procedure

Collect the specimen as usual, using the appropriate collection technique for SurePath®, ThinPrep® or Digene® cervical sampler.

d. Procedural Notes

This test was developed and its performance characteristics determined by the Center for Disease Detection, LLC. It has not been cleared or approved by the USDA.

e. CPT Code(s)

87621

9. HPV, High-Risk DNA, Hybrid Capture II

Test Code(s): [HC HPV]

a. Purpose of Test

HPV is the causative agent of cervical dysplasia and cervical carcinoma. This assay only detects high-risk HPV types: 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59 and 68.

b. Collection Device / Specimen Type

  • i. Digene® specimen transport medium: 1mL
  • ii. Cytyc® ThinPrep® solution: 4.1mL
  • iii. SurePath® CytoRich® fraction: 2mL

c. Collection Procedure

Collect the specimen as usual, using the appropriate collection technique for SurePath®, ThinPrep® or Digene® cervical sampler.

d. Procedural Notes

Do not freeze specimens. Frozen specimens will be rejected for testing.

e. CPT Code(s)

87621

f. Reference Values

HPV HR Negative

10. HPV Genotypes 16 and 18

Test Code(s): [HPV GENO]

a. Purpose of Test

HPV 16 and HPV 18 are estimated to cause 70% of cervical cancers worldwide and that the cumulative risk of (CIN3+) in women with HPV 16 or HPV 18 ranges from 10% to 20%. These two HPV types are thought to be potent carcinogens and should be more effectively targeted in clinical practice.

b. Collection Device / Specimen Type

  • i. Residual PreserveCyt fluid for ThinPrep® samples
  • ii. Residual SurePath® fluid for SurePath® samples

c. Collection Procedure

Collect the specimen as usual, using the appropriate collection technique for SurePath® or Thin Prep®

d. Procedural Notes

  • i. Cervical swabs in Digene HC cervical sampler are not suitable for analysis.
  • ii. Minimum volumes:
  • - ThinPrep® 2mL
  • - SurePath® 1mL

e. CPT Code(s)

HPV GENO 87621

f. Reference Values

HPV Genotypes 16 and 18 Negative

11. Culture, CT (Chlamydia trachomatis)

a. Test Code(s):

[CULT CT]

b. Purpose of Test

C. trachomatis is associated with infections of the mucous membranes of the urogenital system, the upper respiratory tract and the eye. It may be sexually transmitted. Resulting diseases include urethritis, cervicitis, salpingitis, epididymitis, proctitis and lymphogranuloma venereum. Co-infection with C. trachomatis and N. gonorrhoeae can occur. Infections of the upper respiratory tract and eye usually occur in newborns exposed at parturition. In adults, eye infection may be transmitted by hand after contact with secretions. Isolation by tissue culture is recommended when testing individuals under the age of thirteen years.

c. Collection Device / Specimen Type

  • i. V-C-M medium (green cap) tube, or equivalent
  • ii. Dacron® / rayon swab from endocervix, urethra, conjunctiva, rectal mucosa (without feces), fluid aspirate, tissue, nasopharynx or throat.

d. Collection Procedure

  • i. Collect specimen using a Dacron®/rayon swab inserted into a VCM (green) transport medium tube.
  • ii. Label the container with the patient’s name and at least one other patient identifier (e.g. birth date)
  • iii. Completely fill out the Precision Pathology OBGYN Requisition.
  • iv. Put the specimen and requisition in a specimen bag for transportation.
  • v. According to the facility’s courier set-up, call for a pick-up or wait for the scheduled pick-up time
  • vi. Refrigerate the specimen until courier pick up.

e. Procedural Notes

  • i. Wooden shaft and calcium alginate swabs, dry swabs, or swabs in bacterial gel-based transport medium are unacceptable.
  • ii. Refrigerated specimens more than 48 hours old or specimens received at room temperature will be rejected.

f. CPT Code(s)

  • i. Culture, CT 87140
  • ii. Antibody Typing 87110
  • g. Reference Values
    No Chlamydia trachomatis isolated.

12. Culture, Genital

Test Code(s): [CULT GEN]

a. Purpose of Test

The significance of any isolate(s) in pure or mixed culture must be assessed with respect to the source cultured, the organism’s pathogenic portential, the possibility of colonization versus infection, and the number of other organisms recovered from the same culture.

b. Collection Device / Specimen Type

  • i. BD™ blue cap, amies gel medium culture device
  • ii. Vaginal, endocervical, urethral, prostatic secretions

c. Collection Procedure

  • i. Collect the specimen from superficial wounds or skin using BD™ blue cap, amies gel medium culture device.
  • ii. Label the container with the patient’s name and at least one other patient identifier (e.g. birth date)
  • iii. Completely fill out the Precision Pathology OBGYN Requisition.
  • iv. Put the specimen and requisition in a specimen bag for transportation.
  • v. Call for a STAT pick-up. This specimen must be submitted on the day it is collected.

d. Procedural Notes

  • i. This assay does not rule out Neisseria gonorrhoeae (use test “Culture, N. gonorrhoeae”) or Group B Streptococci (use test “Culture, Streptococcus Group B”).
  • ii. Collect the specimen using a culture swab transport device. Indicate the source of the specimen on both the test requisition and the specimen transport device.

e. CPT Code(s)

  • i. Culture 87070
  • ii. Identification 87077
  • iii. Susceptibility
  • f. Reference Values
  • i. Depends on the source
  • ii. Typically “Normal Urogenital Flora” for negative specimens

13. Culture, GC (Neisseria gonorrhoeae)

Test Code(s): [CULT GC]

a. Purpose of Test

Neisseria gonorrhoeae is considered a pathogen whenever isolated and its identification is important in initiating appropriate therapy to prevent the spread of the infection as well as the serious sequelae.

b. Collection Device / Specimen Type

  • i. BD™ blue cap, amies gel medium culture device
  • ii. Urethral, cervical, anorectal or throat

c. Collection Procedure

  • i. Collect specimen using a blue cap, Amies gel transport medium swab delivered within 24 hours of collection.
  • ii. Label the container with the patient’s name and at least one other patient identifier (e.g. birth date)
  • iii. Completely fill out the Precision Pathology OBGYN Requisition.
  • iv. Put the specimen and requisition in a specimen bag for transportation.
  • v. According to the facility’s courier set-up, call for a pick-up or wait for the scheduled pick-up time
  • vi. Maintain the specimen at room temperature – DO NOT REFRIGERATE

d. Procedural Notes

  • i. Includes routine isolation and identification procedures. Susceptibility testing is not routinely performed.
  • ii. Collect specimen using a culture swab transport device. Indicate the source of the specimen on both the test requisition and the specimen transport device.

e. CPT Code(s)

Culture, GC 87081

f. Reference Values

No Neisseria gonorrhoeae isolated.

14. Culture, Trichomonas vaginalis

Test Code(s): [CULT TVAG]

a. Purpose of Test

Trichomonas vaginalis is not part of the normal flora of the genital tract and is considered a pathogen when detected. Its identification is important for the control and treatment of trichomoniasis.

b. Collection Device / Specimen Type

  • i. Trichomonas Inpouch™ culture medium
  • ii. Female – vaginal secretions
  • iii. Male – urethral exudates, prostatic secretions or sediment from centrifuged urine

c. Collection Procedure

  • i. Using a sterile cotton-tipped swab, obtain the specimen and immediately inoculate the culture medium.
    Note: Use only a cotton-tipped swab. The organisms will cling to Dacron or any plastic surface.
  • ii. Close the specimen pouch by rolling over the flap. Secure with the wire strip to prevent leakage.
  • iii. Label the container with the patient’s name and at least one other patient identifier (e.g. birth date)
  • iv. Completely fill out the Precision Pathology OBGYN Requisition.
  • v. Put the specimen and requisition in a specimen bag for transportation.
  • vi. According to the facility’s courier set-up, call for a pick-up or wait for the scheduled pick-up time

d. Procedural Notes

None

e. CPT Code(s)

87081

f. Reference Values

No Trichomonas Seen

15. HIV-2 with Reflex to Western Blot

Test Code(s): XXXXXX

a. Purpose of Test

HIV-2 is closely related to HIV-1 regarding nucleic acid sequence and clinical disease. HIV-2 is endemic to West Africa with nearly all cases in the United States identified in citizens or travelers from West Africa. Rarely, HIV-1 Western blot indeterminate results may be due to HIV-2 infection in a patient who has been exposed to HIV-2. The Immunoblot is useful to confirm repeatedly reactive EIA results.

b. Collection Device / Specimen Type

  • i. Plastic Red Top (no additive) blood collection tube
  • ii. Minimum of 1 mL serum

c. Collection Procedure

  • i. Following the standard phlebotomy venipuncture technique, blood is drawn from a vein, usually from the inside of the elbow.
  • ii. Collect at least 4 mL of whole blood in a plastic red top blood collection tube
  • iii. Label the container with the patient’s name and at least one other patient identifier (e.g. birth date)
  • iv. Completely fill out the Precision Pathology OBGYN Requisition.
  • v. Put the specimen and requisition in a specimen bag for transportation.
  • vi. According to the facility’s courier set-up, call for a pick-up or wait for the scheduled pick-up time

d. Procedural Notes

None

e. CPT Code(s)

86702

f. Reference Values

Non-Reactive

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F. Muscle Biopsy

1.         Call the Laboratory at 210-646-0890 at least 24 hours in advance of the procedure to ensure the Laboratory has a courier on stand-by to pick up the specimen STAT.

Procedures should be scheduled for a no later than 14:00 pick-up time, due to time constraints on specimen processing.

NOTE: If located outside the 1604 loop, muscle biopsy procedures must be scheduled with PPS in advance, so a histotechnician can be on-site for immediate specimen handling.

2.         Select a muscle that is neither asymptomatic, nor end stage.

3.         For proximal weakness, biceps or quadriceps are preferred.

4.         For distal weakness, brachioradialis or anterior tibial muscle tissue is preferred.

5.         Never biopsy the gastrocnemiusmuscle in children.  Gastrocnemius biopsies are only acceptable in adults.

6.         Do not biopsy a muscle recently needled by electromyography or injected with medication.

7.         Infiltrate the skin with anesthesia, but do not infiltratethe muscle tissue.

8.         Incise the skin and muscle fascia.  Retract the skin,exposing the muscle tissue.

9.         As atraumatically as possible, by blunt dissection, longitudinally separate several fascicles (at least 5mm across) to greater than the width of the muscle biopsy clamp.

10.       Insert the muscle biopsy clamp onto the muscle.  Close the clamp tight enough to hold the muscle in place, to prevent contraction.

11.       Incise the muscle outside the edges ofthe clamp, leaving at least 1mm on each edge outside the clamp.

12.       Keep the specimen in the muscle clamps.

13.       The specimen should be put into a small, pre-labeled, container with two-three drops of saline.  DO NOT SUBMERGE specimen in saline.

14.       Then, place the smaller container into the larger container and pack ice around the smaller container.  Be sure the smaller container is labeled with two patient identifiers (including full patient name).

15.       Hand the specimen directly to the PPS Courier for delivery to the laboratory STAT,without any delays.

Note:  Both specimen containers must be labeled with two patient identifiers (patient’s full name, and one other identifier, such as date of birth).  Be certain the requisition is completely filled out with all pertinent information.

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G. Nerve Biopsy

  • a. Submit 3cm of sural/posterior tibial nerve (or less of peroneal/other sensory diminishing nerve), sutured at each end and affixed to a longitudinally-split portion of tongue blade with a notch at each end, so the nerve is lying straight.
  • b. Do not stretch or freeze nerve. Results will be very suboptimal.
  • c. Call PPS at 210-646-0890 for a STAT pick up. Procedures should be scheduled for a no later than 14:00 pick-up time, due to time constraints on specimen processing.
    NOTE: If located outside the 1604 loop, muscle biopsy procedures must be scheduled with PPS in advance, so a histotechnician can be on-site for immediate specimen handling.
  • d. The specimen container must be labeled with two patient identifiers (patient’s full name, and one other identifier, such as date of birth). Be certain the requisition is completely filled out with all pertinent information.

(See procedure 110 for more details.)

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H. Epidermal Small Nerve Fiber Density Testing

Epidermal nerve fiber density testing (ENFD) is a neuropathology test in which a punch skin biopsy is taken to identify small fiber neuropathies by analyzing the nerve fibers of the epidermis.
  • 1. Materials Required
    • a. Acu*Punch Kit
      • i. 1 – Acu*Punch 3mm
      • ii. 1 – Acu*Scissors Curved 3½”
      • iii. 1 – Fine-tip Forceps
      • iv. 2 – Gauze Pads
      • v. 1 – Alcohol Prep Pad
      • vi. 1 – Povidone-Iodine Prep Pad
      • vii. 1 – Oval Bandage
      • viii. Each kit is wrapped in a disposable drape
    • b. 1 – 20mL container of 10% NBF
  • 2. Cutaneous samples must be obtained according to the following protocol, or the specimen will likely be rejected for specimen processing.
  • 3. Specimens must be collected from one or more of the following sites:
    • a. 10cm above the lateral maleolus
    • b. 7cm above the knee
    • c. 7cm below the hip
  • 4. Procedure
    • a. Measure to the intended site (e.g. 10cm above the lateral maleolus) and mark the location with Sharpie.
    • b. Swab the area with the Alcohol Prep Pad
    • c. Infiltrate 1cc of anesthesia in a V-shaped pattern, with the apex of the V proximal to the marked site and the limbs of the V on either side of the Sharpie mark.
    • d. Take the Povidone-Iodine Prep Pad and massage the anesthesia into the injected area. Allow approximately 30 seconds for the anesthesia to set.
    • e. Take the 3mm punch in thumb, forefinger and middle finger and rest it against the site. Gently rotate the punch to incise through the skin. Do not press straight down with the punch, as this will damage the intended specimen. Stop when the metal tip of the punch is about halfway into the skin.
    • f. To remove the specimen, press down on the epidermis on either side of the punch to allow a grip on the side of the specimen. Do not allow forceps-contact with the epidermis directly to avoid evidence of crush artifact.
    • g. Lift the specimen, and freen the base of the punch with the provided Acu-Scissors.
    • h. Place the specimen into the 20mL container of 10% NBF and ensure that it is completely submerged and not allowed to float on the air-fluid interface. Make sure the container is labeled with the patient’s full name and date of birth.
    • i. Place the specimen in a biohazard transport bag with a completed pathology requisition form .
    • j. According to the facility’s courier set-up, call for a pick-up or wait for the scheduled pick-up time

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I. Hirschsprung’s Disease Specimens

Expected Routine TAT 24-48 hours

1. Materials Needed

  • a. Telfa Pad
  • b. Saline
  • c. Sterile container

2. Procedure

  • a. Place the fresh specimen on a “taco” of telfa pad that has been soaked in saline.
  • b. Place the telfa taco into a sterile container. Do not add fixative. A container emptied of fixative is not suitable for this specimen.
  • c. Make sure the specimen container is labeled with two, unique patient identifiers. (e.g. Patient’s full name, and date of birth)
  • d. Put the container on ice.
  • e. Call for a STAT pick-up: 210-646-0890

Note: The laboratory can only accept specimens for Hirshsprung’s between 0300 and 1530, Monday through Friday. Call the Laboratory with any questions.

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J. Urine Specimen Preparation for Microbiology Cultures

See Microbiology section below

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K. Specimen Preparation for Gout

Expected Routine TAT 24-48 hours
In order to maximize accurate results for Pathology, the person collecting and handling the original specimen should:

1. Pre-label a 95% ETOH (alcohol) Container with the Patient’s Name, Collection Date, and Collection Site.

  • a. This specimen MUST NOT BE SUBMITTED INTO FORMALIN.
  • b. This specimen MAY be SUBMITTED FRESH.

2. Insert the specimen into the container. Close the container and tighten the lid.

3. According to the facility’s courier set-up, call for a pick-up or wait for the scheduled pick-up time.

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L. Specimen Preparation for Chromosome Analysis

Expected TAT 2-3 Weeks
In order to maximize accurate results for Chromosome Analysis, the person collecting and handling the original Product(s) of Conception (POC) specimen should:

Pre-label an empty dry biopsy container with the Patient’s Name, Collection Date, Collection Time and Collection Site.

Monday-Thursday

  • Moisten a piece of gauze with saline.
  • Wrap the piece of gauze very loosely around the specimen.
  • Insert the specimen into the specimen container.
  • Seal the container and insert it into a specimen transport bag.
  • Fill out a Patient Surgical Requisition and insert it into the side pocket of the specimen transport bag.
  • Hold the specimen in a 4° Celsius refrigerator until the courier arrives for pick-up.
  • In-town: Call Precision Pathology at 210-646-0890 for a “STAT” pick-up and let them know you have a FRESH Specimen for Cytogenetics.
  • Rural: If the normal courier pick-up time is missed, hold the specimen in a 4° Celsius refrigerator until the next day pick-up.
  • This specimen MUST NOT BE SUBMITTED IN FORMALIN.

Friday

  • Place the specimen in a dry specimen container.
  • Cover the specimen in RPMI.
  • Seal the container and insert it into a specimen transport bag.
  • Fill out a Patient Surgical Requisition and insert it into the side pocket of the specimen transport bag.
  • Hold the specimen in a 4° Celsius refrigerator until the courier arrives for pick-up.
  • Call the Laboratory before 1700 to arrange handling. If the specimen is ready after 1700, call the Pathologist on-call.

Saturday-Sunday

  • Place the specimen in a dry specimen container.
  • Cover the specimen in RPMI.
  • Seal the container and insert it into a specimen transport bag.
  • Fill out a Patient Surgical Requisition and insert it into the side pocket of the specimen transport bag.
  • Hold the specimen in a 4° Celsius refrigerator until the courier arrives for pick-up on Monday. Call and alert the Laboratory to ensure timely pick-up.
  • Reminder: These specimens are to be kept moist with saline-soaked gauze.
    DO NOT SUBMERGE IN SALINE.

    * Products of Conception/Fetal Tissue for general histology processing can be submitted in formalin. Hold for courier pick-up.
    * Fetal Remains for Autopsy must be kept fresh. See procedure 176.100 for more details.

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M. Specimen Preparation for Flow Cytometry

Expected TAT 24-48 hours
In order to maximize accurate results for Flow Cytometry, the person collecting and handling the original specimen should:
1. Pre-label a container of RPMI with the Patient’s Name, Collection Date, Collection Time, and Collection Site.
2. Insert the specimen into the pre-labeled container.
3. Seal the container and insert it into a specimen transport bag with a cold (not frozen) pack.
4. Fill out a Patient Surgical Requisition and insert it into the side pocket of the specimen transport bag.
5. Hold the specimen in a 4° Celsius refrigerator until the courier arrives for pick-up.
6. Call Precision Pathology at 210-646-0890 for a “STAT” pick-up and let them know you have a FRESH Specimen in RPMI for Flowcytometry.
*This specimen MUST NOT BE SUBMITTED INTO FORMALIN.

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N. Specimen Preparation for Bone Marrow

Expected TAT: 24-172 hours (depending on testing needed)

The Clinician will receive a Preliminary Diagnosis call from the Pathologist within 24 hours of specimen receipt.

Contents:

  • 2 bottles Bouin’s Fixative
  • 1 Cold Pack
  • 2 slide containers, each containing 5 slides
  • 1 Specimen Bag
  • 1 Purple Top containing EDTA
  • 1 Instruction Sheet
  • 1 Green Top containing Sodium Heparin1 Transport Box
1. A Bone Marrow study usually consists of an aspirate and biopsy, or just an aspirate.

  • Always be specific about the sampling site (right or left posterior iliac crest or sternum).

2. The optimal amount of aspirate is 4-5 mL. A portion of the aspiration is used at the bedside to make 4-6 particle crush smears.

  • a. To create a particle crush smear, particles should be placed in the center of the glass slide.
  • b. Another slide is positioned directly over the first slide, and thetwo slides are gently placed together and then pulled apart in a planeparallel to their surfaces.
  • i. These smears should be quickly air-dried.
  • ii. Direct smears of the aspirate prior to clotting are also helpful.These are made as you would a Peripheral Blood Smear (see below).
  • c. At least 1-2 mL of aspirate is to be placed in the purple top (EDTA)tube and at least 0.5 – 1mL of aspirate should be placed in the greentop (sodium heparin) tube.
  • d. The remainder of the aspirate is allowed to clot and then is placed in one of the containers of Bouin’s Fixative.

3. The trephine biopsy should immediately be gently imprinted(touched) several times per slide on two or three glass slides to maketrephine imprints. Air dry these immediately. Place the biopsy in thesecond container of Bouin’s fixative.

4. A copy of the CBC and one or two unstained peripheral blood smears ortube of peripheral blood should be accompany the bone marrow. If noperipheral blood was obtained, finger stick smears are acceptable.

5. Special Studies that will be performed as necessary on the marrow are the following:

  • a. Chromosome (Cytogenetic) Studies: Performed on 0.5 – 1.0 mL ofunclotted aspirate in a sodium heparin tube. Maintain at roomtemperature. Performed at CorePath Laboratories on 2020 Babcock, Ste 30SA, TX 78229.
  • b. Flow Cytometry: Place at least 1-2mL of marrow aspirate inEDTA tube. Maintain at room temperature. Performed at CorePathLaboratories on 2020 Babcock, Ste 30 SA, TX 78229.
  • c. FISH: FISH may be performed on the same sample (sod hep tube)sent for chromosome studies. Performed at CorePath Laboratories on 2020Babcock, Ste 30 SA, TX 78229.
  • d. Molecular Diagnostics: Includes polymerase chain reaction(PCR) and Southern blot analysis. These may be performed on the same(EDTA) tube sent for Flow Cytometry. Performed at Clarient on 31Columbia Aaliso, Aiso Viejo, CA 92656.
  • e. Cultures: A second aspirate is required for cultures, in order to maintain sterility. Aspirate about 5mLs into syringe, remove needle, and cap. Usual studiesdone are aerobic/anaerobic, acid fast, and fungal cultures.

6. When the marrow is prepared for transport:

  • a. Be sure ALL SLIDES AND CONTAINERS ARE LABELED WITH THE PATIENT’S NAME and COLLECTION DATE. Indicate the date and time the clot and biopsy were placed in fixative on the requisition.
  • b. Be sure to close containers tightly to omit any specimen leaks, or specimen destruction. The fumes of the Bouin’s fixative have the ability to destroy the slide specimens.
  • c. Place the dried slides into the plastic slide containers for protection. The slides must be thoroughly dry before placing in the plastic containers to prevent artifact.
  • d. Place the bottles of Bouin’s fixative into the provided plastic bag. Seal the bag to eliminate any fumes from escaping. Insert all items into the Bone Marrow Kit Box.

7. Enclosed paperwork should include: The most recent CBCresults, Patient Demographic information, Hematopathology Requisitioncontaining; Patient’s age and/or date of birth, Diagnosis (if known),site of the Biopsy, Requesting physician names, and any Special Studiesthat may be requested. Always Print when filling out specimen requisitions,and do not use colored inks or pencils.

8. Questions? Please call Dr. Peggy Stoll direct at 210-757-8399 or at Precision Pathology Services at 210-646-0890.

9. Send to: Please call Precision Pathology Services to arrange a specimen pick-up.

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O. Specimen Preparation for Microbiology

To report medically-relevant results, the Specimens sent by Clients and received into the Microbiology Laboratory must be properly collected. The Microbiologist has little control over the quality of specimens received. Bearing this in mind, it is imperative to assist Clients with specimen collection questions. The List of Samples for Collection lists sample collection guidelines for Clients and should be utilized as a very important reference. It is to be available to all Microbiology Laboratory Personnel.

1. All samples received for processing must adhere to the following basic criteria:

  • a. Utilize Sterile Containers.
  • b. Label specimen containers properly with specific sources for wounds and type of collection for urines. (i.e. decubitis left great toe; ear, skin lesion left forearm; cath; etc.)
  • c. Utilize proper Transport Media for culture type. (i.e. culturette for routine aerobic wounds; BD culturette transport for urogenital sources; anaerobic transport for anaerobic samples)
  • d. Transport in the proper temperature environment.

2. Specimens recommended for General Microbiology testing:

  • a. Deep or post-operative exudates and drainages for wounds.
  • b. Clean catch, midstream urines – First morning void preferred, catheterized (avoid random urine collections)
  • c. Venous or arterial for Catheter
  • d. Suprapubic Aspirate
  • e. Sputum (lower respiratory secretions) for Respiratory – First morning preferred or aspirated sputum
  • f. Pharyngeal Swab (Vigorously swab tonsillar area, NOT the tongue)
  • g. Nasopharyngeal Swab
  • h. Bronchial Wash
  • i. Transtracheal Aspiration
  • j. Fine Needle Aspiration
  • k. Ear (Otitis Media)
  • l. Eye (Lacrimal duct, corneal scrapings)
  • m. Skin – Discard surface debris and clean around the lesion with a skin disinfectant. Collect a Swab or Aspirate from inside the lesion
  • n. Body fluids – These are normally sterile fluids. Collect aspirates using an aseptic technique.
  • o. Abscesses – Purulent aspirate plus sample from the side wall of the abscess
  • p. Genitourinary Tract – Use a Swab in the approved Transport Media.

3. Specimens not recommended for General Microbiology testing:

  • a. Urines that are very dilute, with low specific gravity.
  • b. Urines that are not transferred to a BD Urine Vacutainer™.
  • c. Urines that are not refrigerated, if delayed in sample receipt and processing.
  • d. Sputum mixed with an abundance of saliva, as evidenced by the Gram’s Stain evaluation
  • e. Stool samples that have been refrigerated (Shigella and some Salmonella do not survive).
  • f. Stool samples that are not placed in preservative, when there is a delay in reaching the Laboratory – (pathogens are overgrown by normal flora)

4. Anaerobic Specimen(s)

  • a. Preferred Anaerobic Specimen(s):
  • i. Abscess or deep wound, tissue, normally sterile body fluids other than blood
  • ii. Transtracheal aspirations
  • iii. Bronchoscopic specimens obtained by the Double Lumen technique
  • iv. Suprapubic urine aspirates obtained from bladder, nephrostomy tube or suprapubic catheterization
    • Genital specimens from these sites only
    • Cul-de-sac aspiration
    • Culdocentesis
    • Transfundal aspiration
    • Placenta
    • Fallopian tube
    • Intrauterine devices
    • Surgical specimens
  • v. Exudates
  • vi. Aspirated pus from deep wound or abscesses
  • vii. Tympanocentesis fluid

5. Rejection Criteria for Anaerobic Specimen(s):

  • a. Specimens received in expired media or frozen
  • b. Specimens not submitted in anaerobic transport media/device
  • c. Specimens collected from the following sites:
    • i. Superficial wounds
    • ii. Throat
    • iii. Nasopharyngeal
    • iv. Feces/rectal swabs
    • v. Urine (catheter or voided)
    • vi. Vaginal
    • vii. Cervical
    • viii. External ear
    • ix. External eye
    • x. Sputum/Bronchoscopic specimens not collected by Double Lumen technique

6. Specific Collection Guidelines

  • a. Use a sterile, appropriate collection media. Ensure that the collection media has not exceeded the expiration date.
  • b. Label the collection media with at least two Patient Identifiers. Always submit all specimens with the patient’s full name and DOB. Other acceptable patient identifiers are Social Security Number and/or Medical Record Number.
  • c. Record the date and time collected on the requisition or the specimen container.
  • d. Collect the specimen prior to initiating antibiotic therapy.
  • e. To prevent deterioration of pathogens, deliver all specimens in a timely manner. It is recommended that collection time to setup time for all specimens is less than 24 hours.
  • f. Urines collected in a BD Vacutainer™ can be refrigerated for up to 2 days, but should be delivered for processing within 48 hours of collection.
  • g. Refrigeration can deteriorate or inhibit growth of some pathogens. Collection swabs should be at room temperature at all times and submitted within 48 hours of collection.
  • h. Always specify on the requisition the specimen collection site. Specify as right or left side if applicable.

7. Smears

  • a. Some Clinicians may choose to recover exudates for Gram Stain immediately at the time of surgical or wound management procedures.
  • b. Label the frosted end of the slide with the Patient’s name and a second Patient Identifier. Smear a thin layer of specimen on the slide and allow the slide to thoroughly air-dry. Collate this slide with any other corresponding culture specimens and prepare for transport to the Laboratory in a labeled slide holder.

NOTES:

* If the individual slides are not labeled, the specimen will be returned to the Clinician for labeling. This is compliant with CAP and CLIA guidelines on specimen labeling.

* DO NOT use slide holders for any specimens other than smears, as these containers are not sterile. Be sure to allow any slides to dry before placing them in the transport slide container.

8. Specific Use of Collection Equipment

  • a. BD BBL™ CultureSwab™ (RED TOP)
    Note: Red tops are preferred for the recovery of aerobic organisms.
    • i. Ensure that the collection swab has not exceeded the expiration date.
    • ii. Remove the swab from the packaging and take care to employ aseptic technique. Aseptic technique means avoiding contact with work surfaces to avoid contamination of the swab.
    • iii. Discard the top cap of the collection tube.
    • iv. Collect the specimen and insert the swab into the collection tube immediately.
    • v. Swabs should be touching the transport media on the bottom of the collection tube.
    • vi. Maintain the culture tube at room temperature throughout transport interval and submit within forty-eight (48) hours of collection.
    • vii. Before submitting the specimen, ensure the specific collection guidelines in Section 6 have been followed.
  • b. BD BBL™ Culture Swab™ Plus (BLUE TOP):
    Note: Blue tops are optimal for recovery of both anaerobic and aerobic organisms.
    • i. Ensure that the collection swab has not exceeded expiration date.
    • ii. Remove swab from package and take care to employ aseptic technique. Aseptic technique means avoiding contamination of the swab coming into contact with work surfaces.
    • iii. Discard top cap of collection tube.
    • iv. Collect specimen and place swab into the gel transport medium. Make certain swab is submerged in the gel transport media for better organism recovery.
    • v. Maintain at room temperature throughout transport interval and submit within 48 hours of collection.
    • vi. Before submitting specimen, ensure the specific collection guidelines in section 3.3 have been followed.
  • c. BD Viral Transport for Viruses, Chlamydiae, Mycoplasmas and Ureaplasmas:
    • i. Ensure that the collection container has not exceeded expiration date.
    • ii. Peel apart plastic packaging and discard.
    • iii. Remove and utilize one of the swabs provided for aseptic collection of sample. The smaller rod-like swab should only be used for chancres or genital ulcers on male patients.
    • iv. After collection of specimen, remove red cap from viral transport tube and insert swab. Submerge the swab into the media and break off swab shaft by bending the shaft against the rim of the tube.
    • v. Recap the tube securely.
    • vi. More than one tube may be submitted if more than one type of viral culture is requested to ensure adequate specimen for culture set-ups.
    • vii. Transport immediately to the lab at 2-30°C within 24 hours.
    • viii. Before submitting specimen, ensure the specific collection guidelines in section 3.3 have been followed.
  • d. BD Vacutainer® Urine Transport:
    • i. Ensure that the collection container has not exceeded expiration date.
    • ii. First collect urine specimen according to clean catch procedure for male or female.
    • iii. Remove the straw and vacutainer tube from the package using care not to contaminate this collection device.
    • iv. Do not place on workbench. This will contaminate the collection device.
    • v. Immediately upon removal from the package insert the tip of straw transfer device into the urine specimen cup and insert the Vacutainer tube in holder portion of straw device.
    • vi. Push the vacutainer all the way into the holder. The vacuum of the tube will aspirate the urine into the collection device.
    • vii. Transfer device should be discarded in sharps container after use.
    • viii. Shake tube vigorously after collection.
    • ix. Refrigerate if not immediately transported to lab. Submit refrigerated specimen no later than 48 hours after collection.
    • x. Before submitting specimen, ensure the specific collection guidelines in section 3.3 have been followed.
  • e. Remel Agar and Media
    • i. Ensure that the collection media has not exceeded expiration date.
    • ii. The following media should be kept refrigerated when not in use:
      • Sabouraud Dextrose plates and slants
      • Chocolate plates
      • Blood/TSA plates
      • Thioglycollate Broth
    • iii. Allow plates/media to warm up to room temperature prior to use.
    • iv. Plates should be inoculated, streaked for colony isolation and taped closed.
    • v. Transport plates immediately with lid face down and agar side up, at room temperature.
    • vi. Before submitting specimen, ensure the specific collection guidelines in section 3.3 have been followed.
  • f. Medscand Sample Collection Kits for use with The ThinPrep® Pap Test for HPV (Human Papilloma Virus)
    • i. Ensure that the collection media has not exceeded expiration date.
    • ii. Refer to Medscand Sample Collection Kit instructions for The ThinPrep® Pap Test specific collection guidelines on exo/endocervix and rectal specimens.
    • iii. Transport specimen at room temperature.
    • iv. Before submitting specimen, ensure the specific collection guidelines in Section 6 have been followed
  • g. Sterile Tubes
    • i. Utilized for placing various specimens needed for culture, (i.e. toenail clippings, skin scrapings, and small tissues).
    • ii. Specimens submitted in unsterile containers for culture are not suitable, and will be rejected.
    • iii. Transport at room temperature within 24 hours after collection.
    • iv. Before submitting specimen, ensure the specific collection guidelines in section 3.3 have been followed.
  • h. Sterile Saline Tubes
    • i. Used for Acanthamoeba (Culture)
    • ii. Preferred specimens: Eye Wash; Corneal Biopsy; Contact Lens/Fluid; cerebral; lung-collected aseptically
    • iii. Transport at room temperature within 24 hours after collection.
    • iv. Before submitting specimen, ensure the specific collection guidelines in section 3.3 have been followed.

9. Unacceptable Specimen Criteria:

  • a. Excessive time between collection of specimens and delivery to the laboratory. Most specimens have a 24 hour transport limit. Urines should be held for no more than 48 hours if refrigerated. This practice of holding urines for this extended time is not recommended and can cause great discomfort to the patient since it will delay treatment of bladder and urinary tract infections.
  • b. Improper Labeling: If specimen is missing two patient identifiers or requisition and specimen label are discrepant or incomplete, the collection staff will be asked to correct.
  • c. Dry Culturette swab: Transport media ampule unbroken
  • d. Grossly leaking or contaminated specimen container: Biologic hazard
  • e. Inappropriate specimens including:
    • i. Specimens for anaerobic culture containing aerobic normal flora (e.g. mouth, skin, vagina, bowel).
    • ii. Urinary tract catheter or tip.
    • iii. Gastric contents for bacteriology (except neonates)
    • iv. Samples received in a syringe with the needle still attached.
    • v. Specimens received in non-sterile containers.
    • vi. Stools for ova and parasites from patients who have received certain exogenous substances.
    • vii. Insufficient quantity of specimen received for tests requested.
  • f. If a specimen is found to be unacceptable, testing will not be performed and the collection staff will be notified. Documentation in the specimen return log will be performed for all unacceptable specimens and will be monitored on the pre-analytical action report on a monthly basis.

10. Safety:

  • a. Collect and transport all specimens in keeping with the type of isolation in use at the time of sampling.
  • b. Transport all specimen types to lab in appropriate biohazard transport bag.
  • c. Make sure the cap or lid is secure on each specimen transport device to prevent possible leakage or spillage.
  • d. If a specimen leaks, dispose of according to your institutions’ implemented biohazard waste disposal procedures and recollect specimen.
  • e. If a specimen is dropped so that the contents leak onto an environmental surface:
    • i. Put on a pair of gloves.
    • ii. Handle the specimen as contaminated trash
    • iii. Clean the affected surface with 10% bleach or disinfectant wipe.
    • iv. Recollect specimen

11. Transport Conditions: Please refer to section 3.5 “Specific Use of Collection Equipment” for individual transport conditions.

12. Maximum transport time is 24 hours to 48 hours. Specimens held up to 72 hours such as weekend collection contingency will be processed but a disclaimer will be issued for sub-optimal recovery conditions.

13. Your marketing representative will provide detailed collection instructions and any changes or problems to collection methods. The marketing representative will also provide contact numbers for pick-up, collection media, requisitions and lock boxes upon request.

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P. Requisitions

Attached to the properly fixed specimen should be a completed Request for pathology services requisition. The requisition form should provide information as needed for a correct interpretation of the histologic specimen, including the pre- and post- operative diagnosis and clinical data.

ALWAYS PRINT when filling out the Specimen Requisitions.

1. Precision Pathology has four types of Requisitions.

  • Surgical Pathology Requisition
  • Hematopathology Requisition
  • Gastroenterology Requisition
  • OBGYN Requisition

2. Required Sections of the Requisition:

  • a. Patient Information Sections
    • Patient Name
    • Date of Birth
    • Social Security Number
    • Account Number
    • Sex
    • Age
    • Insurance Documentation (may be attached to Requisition, please indicate)
  • b. ICD-9 Code
  • c. Specimen Information section
    • Collection Date
    • Collection Time
  • d. Requesting Physician section
    • Mark the patient’s specimen-collecting Physician along with any copy-to Physician(s).
  • e. Pre-Operative Diagnosis
    • Fill in the Pre-op Diagnosis section.  Patient History can be included in this section.
  • f. Therapy Section
  • The laboratory does not have access to the Patient’s Medical Record.  Please include the following information to help assist in giving the most accurate diagnosis for the Patient:

    • Did the Patient have surgery?
    • Is this a routine test?
    • Did the patient have radiation or chemotherapy?
    • Did the patient have abnormal biopsies in the past?
    • Does the patient have a history of cancer?
    • For Bone Marrow patients, please include any blood testing results.

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Q. Transportation

Methodology:

  • 1. Specimens should be kept and stored in the clinic’s designated area to await transportation by courier or overnight mail to the Laboratory for processing.
  • 2. All specimens should be transported inside of a designated rigid specimen transport container within the vehicle. Specimens should not be transported loosely inside the courier vehicle.
  • 3. Specimens are not to be transported in extreme temperatures and all measures should be taken to maintain specimen viability.
  • 4. In case of an accident while transporting biohazards, Precision Pathology Services is to be called immediately and the Medical Director is to be notified.
  • 5. Transportation occurrences which affect specimen quality should be reported to the Laboratory Manager immediately. The occurrence will be investigated and documented. All transport occurrences require QI action.

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R. Radioactive Material

Precision Pathology Services DOES NOT ACCEPT radioactive material; which may include Sentinel Lymph Nodes, Breast Biopsies, Prostate “Seeds” etc. 
Note: During routine facility visits, if requested, pathologists will remove radioactive seeds from tissue specimens and give the seeds to the facility for storage or disposal. The tissue will then be submitted to PPS by the facility per routine protocols.

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S. FRESH Tissue that is Positive or Suspicious for HEPATITIS B, C, HIV, or TUBERCULOSIS:

Precision Pathology Services DOES NOT ACCEPT fresh Hepatitis B, C, HIV, or TB specimens. Due to exposure risk, these specimens must be submitted to the lab in 10% Neutral Buffered Formalin or Bouins fixative. Peripheral Blood smears need to be performed at the Client’s site on these specimens.

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T. Creutzfeldt-Jakob Disease

The lab does not accept any specimen suspected of Creutzfeldt-Jakob disease, include brain or cerebral spinal fluid.

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U. Request for Body Part Returned

Patients requesting body parts to be returned may be done by following the Texas Administrative Code Title 25 Part 1, Subchapter K.
Human materials removed during surgery, labor and delivery, autopsy, embalming, or biopsy shall be subjected to proper disposal method per Tex Admin Code Title 25. (See reference attached).
Specimens may be returned to patient for proper burial by signing release forms (see AUTHORIZATION FOR BODY PART RELEASE attached) and by scheduling with funeral home to retrieve such requested Part. Funeral Home and transportation fees are patient’s responsibility.

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V. Request for Fetus Release

Parents requesting fetus to be returned may be done by following the Texas Administrative Code Title 25 Part 1, Subchapter K.
Human materials removed during surgery, labor and delivery, autopsy, embalming, or biopsy shall be subjected to proper disposal method per Tex Admin Code Title 25. (See reference attached).

Fetus may be returned to parents for proper burial by signing release forms (see AUTHORIZATION FOR FETUS RELEASE attached) and by scheduling with funeral home to retrieve such requested Part. Funeral Home and transportation fees are patient’s responsibility.

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W. Request for Returns (General)

Any biohazardous substance collected and sent for pathology must follow the Texas Admin Code Title 25 Part 1, Subchapter K.  e.g.  Hardware, gallstones, etc

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X. Requests for Technical-Only Aspects

1. Microscope Slide Staining

  • a. Tissue Fixative should be Neutral Buffered Formalin.  If any other fixative type is used, call the lab to inquire if staining will work.
  • b. Slides are to be positive-charged.
  • c. Sections are to be picked up on positive-charged slides from a Distilled Water Bath
  • d. Sections are to be placed 15mm below the frosted cover and at least 5mm from the bottom edge of the slide.  Do not let the tissue go closer than 5mm from the bottom edge of the slide.
  • e. Slidse are to container two unique identifiers.
  • f. Do not dry slides in the oven.  Send the slides to the laboratory directly from the water bath.
  • g. For each stain requested, send four slides.

2. Specimen Processing

  • a. Follow the Routine Specimen Collection instructions.
  • b. Notate on the Requisition in RED the technical aspect being requested.
    • Processing
    • Process and Embed
    • Process, Embed, Microtomy – list the number of slides and how many sections on each slide
    • Process, Embed, Microtomy, staining – list the requested stains
    • Other – Be very specific.  Call the laboratory to consult with a histotechnician if needed.

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References

Carson, F.L., Histotechnology A Self-Instructional Text, 2nd ed., ASCP Press, Chicago, 1997, pp. 80-111.

Sheehan, D.C. and Hraphak, B.B.: Theory and Practice of Histotechnology, 2nd ed., 1980, Mosby Company, St. Louis.

Gomella, L.G. and Haist, S.A.: Clinician’s Pocket Reference, 2002, McGraw Hill Company inc., United States of America.

CAP Checklist, June 2010 GEN.40050, GEN.40100, GEN.40104, GEN.40108, GEN.40116, GEN.40125, GEN.40491, GEN.40500, GEN.40511, GEN.40515, GEN.40522, GEN.40700

CLIA Regulation, 1989. 

Texas Administrative Code Title 25, Part 1, Subchapter K (See attached to this Collection Manual).Precision Laboratories Collection Manual, 2010

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