954. Cytologic Validation of Unique Cervical Cancer Test Bundle Utilizing Target Enriched Multiple HPV E6/E7 DNA Genotyping Assay and mRNA E6/E7 OncoTect™
Session: Poster Abstract Session: Sexually Transmitted Infections
Friday, October 4, 2013
Room: The Moscone Center: Poster Hall C
  • 954_IDWposter.pdf (1.1 MB)
  • Background: The goal of cervical cancer screening is to determine “Who is at risk to develop high-grade dysplasia”.  FDA-approved assays which target the L1 or E1 region of the HPV virion are biased to false negative, particularly in advanced disease.  A genotyping assay detecting the more stable E6/ E7 region of the virus used with an assay identifying the oncogenic activity of the virus in infected cervical cells, improves the specificity of screening, resulting in decreased unnecessary gynecological procedures.

    Methods: 102 liquid based cervical samples were tested for HPV DNA using Diatherix proprietary  target enriched multiplex PCR (Tem-PCR) and by HPV OncoTect™ (incellDX).  Morphologic cytology was performed.  Diatherix Tem-PCR HPV genotyping assay consists of E6/E7 molecular targets for high risk (HR) HPV types, including  16 and 18.  Nested PCR, with tagged, target specific primers providing initial target gene identification, and common flanking Super Primers providing bulk amplification, is used.  A cell control assures specimen quality.  The HPV OncoTect™ assay quantitatively measures over-expression of HPV E6/E7 mRNA in each cervical cell and the total number of over-expressed cells. 

    Results: Cases were sorted by cytologic evaluation.  NILM n=24, ASCUS n=38, LSIL n=13, and HSIL n=27.  HRHPV was detected in 6 NILM, 33 ASCUS, 10 LSIL, and 26 HSIL. mRNA was detected in 4 NILM, three of which were negative for HPV; two ASCUS, one negative for HPV; 6 LSIL, three negative for HPV; and 26 HSIL.  One HSIL was negative for HPV, positive for mRNA. 

    Conclusion: Detection of HRHPV DNA in 26 of 27 HSIL specimens, validates Diatherix Tem-PCR HPV assay compared to the clinical endpoint of HSIL.  The HSIL specimen negative for HPV was mRNA positive suggesting transformation.  There is 100% concordance between cytology and HPV Tem-PCR.   HRHPV were detected in 75 of 102 patients studied, while mRNA was detected in only 12, suggesting that only these 12 were at risk for high-grade dysplasia.  In addition, mRNA was detected in 5 patients in the absence of HRHPV, suggesting that these patients tested with this bundle of tests, were also at risk for high-grade dysplasia. The combination of HPV Tem-PCR and mRNA expression not only defines who is infected but who is infected with transformation.


    Carol Quinter, PhD, Microbiology, Kettering Medical Center, Kettering, OH, Jacqueline Emery, M.D., Professional Pathology 'services, Columbia, SC and Karen Wolters Wolters, MLS (ASCP) MB, Molecular Science, Kettering Health Network, Kettering, OH


    C. Quinter, Diatherix Laboratory: Consultant and Speaker's Bureau, Consulting fee

    J. Emery, None

    K. W. Wolters, Diatherix Laboratories: Employee, Salary

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