Intracellular Human Papillomavirus E6, E7 mRNA Quantification Predicts CIN 2+ in Cervical Biopsies Better Than Papanicolaou Screening for Women Regardless of Age

Abstract

Objectives.-To determine the performance of in-cell HPV E6, E7 mRNA quantification (HPV OncoTect) for the detection of high-grade cervical intraepithelial neoplasia in women younger than 30 years.Design.-We analyzed 3133 cytology specimens from a screening population of women aged 19-75 years investigate HPV OncoTect as a triage/secondary screening test for atypical squamous cells of undetermined significance (ASCUS) and low-grade squamous intraepithelial lesion (LSIL) cytology in women younger than 30 years. Test results were compared to histology in 246 cases.Results.-The sensitivity of E6, E7 mRNA was 89% for CIN 2+ and 100% for CIN 3+ lesions in women 30 years and older. In women younger than 30 years, the sensitivity of E6, E7 mRNA for CIN 2+ lesions was 88% for CIN 2+ and 92% for CIN 3+ lesions. Abnormal cytology ($ASCUS) exhibited a sensitivity of 89% for CIN 2+ and 100% for CIN 3+ in women 30 years and older and 96% sensitivity for CIN 2+ and 93% sensitivity for CIN 3+ in women younger than 30. The specificity of E6, E7 mRNA was .80% for CIN 2+ and CIN 3+ in both groups of women compared to a specificity of abnormal cytology of ,10% for CIN 2+ and CIN 3+ in both groups.Conclusions.-HPV OncoTect demonstrates a performance that would be effective for ASCUS/LSIL triage in women including those younger than 30 years.(Arch Pathol Lab Med. 2012;136:956-960; doi: 10.5858/ arpa.2011-0180-OA) E ffective cervical cancer screening programs like those implemented in the United States have reduced the incidence and the mortality of cervical cancer.1 Current cervical cancer screening recommendations include the use of more sensitive human papilloma virus (HPV) assays as an adjunct to cervical cytology in women more than 30 years old in an attempt to identify those women at risk for high-grade cervical intraepithelial neoplasia (CIN 2+) or cervical cancer lesions. 2,3Women younger than 30 years old were shown to have more transient HPV infections, even by high-risk HPV types, making screening for CIN 2+ in this age group solely dependent on cytologic diagnosis. 4 Because approximately 15% of cervical cancer cases occur in women between 20 and 34 years of age, a need for improved cervical cancer screening exists in this age group. 5 To improve the performance of cervical cancer screening especially in women younger than 30 years, we investigated the use of a quantitative E6, E7 mRNA assay (HPV OncoTect, Incell DX, Menlo Park, California) that determines oncogene overexpression on a cell by cell basis using high throughput flow cytometry. 6,7 The E6 and E7 oncogenes drive cervical cell transformation leading to cancer. The expression of the HPV E6 gene leads to degradation of the tumor suppressor gene p53, and similarly, the HPV E7 gene associates with the tumor suppressor gene pRB.8,9 E7-pRB association results in the upregulation of an E2F-like transcription factor, allowing progression of the cell cycle through the G 1 /S phase. 10 The RB gene product represses expression of cyclin-dependent kinase inhibitor 2A (p16),...

RNA extraction method is crucial for human papillomavirus E6/E7 oncogenes detection

“…Women that had suffered from lesion progression were mostly positive for HPV mRNA (94.4% bioMérieux and 58.82% Roche), while viral expression positivity was found to be lower in women that had remained with the same type of lesion (43.75% bioMérieux and 6.06% Roche). These results agreed with other authors that suggested E6/E7 mRNA expression as a prognostic factor for high-grade lesions and confirmed viral oncogene expression association with severity of cervical lesions [27–29]. …”

High Risk HPV E6/E7 Oncoprotein Expression in Women with High Grade Squamous Intraepithelial Lesion

“…30,31 The sensitivity of the E6/E7 test, 84% for CIN2/3, was greater than that observed by Jeantet et al, 32 who determined a 73.6% sensitivity for CIN2 and 82.2% for CIN3. These numbers were lower than that obtained by Pierry et al, 33 who observed a 89% sensitivity for CIN2 and 100% for CIN3. These numbers led some researchers to suggest the use of the E6/E7 test as a screening test in cases with abnormal cytology or HPV-DNA tests.…”

Effectiveness of a two‐stage strategy with HPV testing followed by visual inspection with acetic acid for cervical cancer screening in a low‐income setting

“…[46][47][48] In conclusion, adding VIA to HPV testing improves PPV of cervical cancer screening, but at the cost of an important loss of sensitivity. Further studies are warranted to explore this strategy.…”

“…Efforts must therefore emphasize on either finding ways to improve VIA accuracy like cervicography or visual inspection with acetic acid and magnification, 44,45 by reviewing criteria for VIA positivity or looking for other triage tools that might be more reliable like VILI, 18,45 or molecular tests such as E6/E7 messenger RNA (mRNA) detection. [46][47][48] In conclusion, adding VIA to HPV testing improves PPV of cervical cancer screening, but at the cost of an important loss of sensitivity. Further studies are warranted to explore this strategy.…”

“…Sociodemographic and reproductive information of the study population and comparison with HPV-positive women and both HPV and VIA positive women are presented in Table 1. Median age was 41 years [interquartile range (IQR), [36][37][38][39][40][41][42][43][44][45][46][47][48][49][50]. Most women were married (58.2%) or widowed (17.5%) and well educated (secondary education or more, 66%).…”