Collection Manual: HPV DNA testing from
ThinPrep™ Vial

Description: Samples for High-risk HPV DNA tests can be performed from a ThinPrep Pap Test collection. Testing is usually performed subsequent to a ThinPrep Pap Test (and from the same vial) though samples may be collected for High-risk HPV DNA testing only. The sampling targets are the ectocervix and endocervix. NOTE: High-risk HPV DNA TESTING can also be performed from the Digene Cervical Sampler with brush (not swab). See instructions for GC/CT collection as outlined in the next section, Collection Procedures 3.2.

ThinPrep Pap kit: PreservCyt™ Solution (CAUTION: POISION, FLAMMABLE), collection devices, requisition form, biohazard transport bag.

1. High-risk HPV DNA tests may be ordered three ways:
a) Case-by-case Reflex Test: When ordering a ThinPrep Pap Test, on the requisition form check “HPV High Risk” then check the conditions for HPV testing (ie, when the result is ASC-US, ASC-H , LSIL or any result)
b) Standing Order Reflex Test: Submit a standing order form to Pathology Laboratory. This will be kept on file and verified annually. You may change or cancel the standing order at any time by submitting another form. Forms are available by calling the laboratory.
• CURRENT ORDER, IRRESPECTIVE OF PAP RESULTS: On the requisition check “HPV High-risk” then “any result”.
• RETROACTIVE ORDER: HPV Testing may be ordered up to 2 weeks after the date of collection. A new requisition must be submitted (see below).
NOTE: If High-risk HPV DNA has been performed within the last 12 months, reflex testing will not be repeated unless specifically requested.

2. Instruct patients to:
• Avoid douches and intercourse 24 hours prior to examination.
• Try to schedule the exam for the non-menstrual portion of her cycle.

3. Complete a requisition form.
White copy to the lab. Lack of data may result in specimen delay or rejection. Include:
• Clinician name, location and date of service.
• Patient demographics: Name, address, phone number, date of birth and social security number.
• Insurance information: copy of insurance card(s) or face sheet.
• Check “HPV High-risk” and specify the criteria for performing the test (ie if ASC-US, ASC-H, LSIL in any combination or, perform High-risk HPV testing regardless of the Pap outcome.
• When ordering High-risk HPV testing retroactively send a requisition to the laboratory (fax, courier or mail) and include clinician name, patient name and birth date, Pap Test accession number, date of request and specify for HPV High-risk testing.

4. Label the vial with the patient’s first and last name.
Collection Procedures 3.1, page 1 of 2

5. Do not apply lubricant to the speculum.
• Moisten the speculum with water. Lubricant will obscure the cells.

6. Visualize and prepare the cervix.
• Do not apply acetic acid or perform other procedures prior to collection of Pap and HPV tests.

7. Collect a cervical sample.
• Gently rotate the plastic spatula 360 around the face of the ectocervix. Include borders of “erosions”. Immediately swirl the spatula 10 times in the vial.

8. Collect an endocervical sample.
• Insert the cytobrush into the endocervical canal until only a few brush bristles remain exposed.
• Slowly rotate ¼ turn. Added rotation is unnecessary and may cause obscuring blood.
• Rinse the brush by rotating it 10 times while pressing it against the inner vial wall. Then swirl the brush vigorously to further release material.

9. Verify that the vial lid is properly seated and tightened.

10. Submit the specimen in a biohazard transport bag.
• Place the vial in the zip-loc portion of the bag and the requisition form in the outer sleeve

11. Refrigeration is not required.
• Samples in PreservCyt are viable for 3 weeks and do not require refrigeration.
• High-risk HPV Testing may be ordered subsequent to ThinPrep Pap results from the same vial if ordered within 2 weeks of the date of collection. If the specimen is between 2 weeks and 3 weeks from the date of collection, call Pathology Laboratory to determine if testing is still viable.

12. Limitations.
• When specimens are submitted for ThinPrep Pap Testing and High-risk HPV Testing, the ThinPrep Pap Test will take precedence. Afterwards, there must be a least 4 ml of sample remaining for High-risk HPV DNA Testing. In about 4% of cases, there will be insufficient material for HPV Testing, in which case a report will be provided explaining that the quantity was insufficient for testing. No charge will be generated for these cases.