Specimen Requirements

Alternate Specimen Requirements

Stability

Days Performed

Mon - Sat

Turnaround Time

5 - 7 days

Methodology

Reference Range

Special Info

In order to facilitate accurate and complete interpretation of cervical screening results, hrHPV test results associated with a cytology report will be held until the cytology report is finalized.

Clinical Info

Persistent infection with human papillomavirus (HPV) is the principal cause of cervical cancer and its precursor cervical intraepithelial neoplasia (CIN). The presence of HPV has been implicated in greater than 99% of cervical cancers worldwide. HPV is a small, non-enveloped, double-stranded DNA virus, with a genome of approximately 8000 nucleotides. There are more than 118 different types of HPV and approximately 40 different HPVs that can infect the human anogenital mucosa. However, only a subset of approximately 14 of these types are considered high-risk for the development of cervical cancer and its precursor lesions. The cobas HPV test performed on the Roche cobas 4800 platform is multiplexed qualitative real-time PCR assay that detects and differentiates between high-risk HPV (hrHPV) types 16, 18, and other (31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 66, and 68). The assay targets a ~200 nucleotide portion of the polymorphic L1 region of the HPV genome, while type-specific probes labeled with different fluorophores differentiates between hrHPV types 16, 18, and other. The test does not detect low-risk HPV types. The assay is FDA-approved for cervical cancer screening in the following situations: reflex testing from ASCUS cervical cytology test results for patients 21 years and older, co-testing with cervical cytology for patients 30 years and older, and first-line primary cervical cancer screening for patients 25 years and older. This lab only accepts specimens collected in ThinPrep PreservCyt solution. This assay has been additionally modified and validated as a lab-developed test to accept fine-needle aspirate (FNA) specimens in ThinPrep PreservCyt solution from patient suspected to have or diagnosed with squamous cell carcinoma of the head and neck. Determination of tumor hrHPV-status in patients suspected to have oropharyngeal squamous cell carcinoma (OPSCC) is critical to staging, prognostication, and therapeutic decision-making. While p16 immunohistochemistry (IHC) is a surrogate marker that can be used to predict hrHPV status, it may be either impossible to perform or unreliable on FNA specimens with scant material. Additionally, p16 IHC can be nonspecifically positive in SCCs from other sites. Accurate classification and staging of a tumor is enhanced when more than one biomarker is utilized (i.e. p16 IHC alongside hrHPV-specific testing) to assess for hrHPV status.

Clinical Limitation

Detection of high-risk HPV is dependent on the number of copies present in the specimen and may be affected by specimen collection methods, patient factors, stage of infection and the presence of interfering substances. Though rare, mutations within the highly conserved regions of the genomic DNA of Human papillomavirus covered by the cobas HPV Test’s primers and/or probes may result in failure to detect the presence of the viral DNA. For FNA specimens from patients suspected of head and neck squamous cell carcinoma, hrHPV PCR results should be evaluated in the context of clinical history and cytology/pathology results (i.e. tumor type and location, p16 immunohistochemical stain, and hrHPV in-situ hybridization studies if available). Assay analytical sensitivity may be diminished with suboptimal collection or specimen pre-processing. Visibly bloody specimens and those contaminated with excessive ultrasound gel may cause assay inhibition leading to false-negative or indeterminate results. If results are incongruent with history, recommend submitting a dedicated specimen for re-testing or request an alternate method for hrHPV detection.

Clinical Reference

1. Fontham ETH, Wolf AMD, Church TR, Etzioni R, Flowers CR, Herzig A, Guerra CE, Oeffinger KC, Shih YT, Walter LC, Kim JJ, Andrews KS, DeSantis CE, Fedewa SA, Manassaram-Baptiste D, Saslow D, Wender RC, Smith RA. Cervical cancer screening for individuals at average risk: 2020 guideline update from the American Cancer Society. CA Cancer J Clin. 2020 Sep;70(5):321-346. doi: 10.3322/caac.21628. Epub 2020 Jul 30. PMID: 32729638. 2. Perkins RB, Guido RS, Castle PE, Chelmow D, Einstein MH, Garcia F, Huh WK, Kim JJ, Moscicki AB, Nayar R, Saraiya M, Sawaya GF, Wentzensen N, Schiffman M; 2019 ASCCP Risk-Based Management Consensus Guidelines Committee. 2019 ASCCP Risk-Based Management Consensus Guidelines for Abnormal Cervical Cancer Screening Tests and Cancer Precursors. J Low Genit Tract Dis. 2020 Apr;24(2):102-131. doi: 10.1097/LGT.0000000000000525. Erratum in: J Low Genit Tract Dis. 2020 Oct;24(4):427. PMID: 32243307 3. American College of Obstetricians and Gynecologists. Updated guidelines for management of cervical cancer screening abnormalities. Practice Advisory. Washington, DC: American College of Obstetricians and Gynecologists; 2020. Available at: https://www.acog.org/clinical/clinical-guidance/practice-advisory/articles/2020/10/updated-guidelines-for-management-of-cervical-cancer-screening-abnormalities. Retrieved April 12, 2021. 4. Maghami E, Ismaila N, Alvarez A, Chernock R, Duvvuri U, Geiger J, Gross N, Haughey B, Paul D, Rodriguez C, Sher D, Stambuk HE, Waldron J, Witek M, Caudell J. Diagnosis and Management of Squamous Cell Carcinoma of Unknown Primary in the Head and Neck: ASCO Guideline. J Clin Oncol. 2020 Aug 1;38(22):2570-2596. doi: 10.1200/JCO.20.00275. Epub 2020 Apr 23. PMID: 32324430. 5. Baldassarri R, Aronberg R, Levi AW, Yarbrough WG, Kowalski D, Chhieng D. Detection and genotype of high-risk human papillomavirus in fine-needle aspirates of patients with metastatic squamous cell carcinoma is helpful in determining tumor origin. Am J Clin Pathol. 2015 May;143(5):694-700. doi: 10.1309/AJCPCZA4PSZCFHQ4. PMID: 25873503.