Prospective evaluation of E6/E7 mRNA detection by the NucliSENS Easy Q HPV assay in a stepwise protocol

Abstract

The objective of the study was to evaluate prospectively the added value of E6/E7 mRNA detection in a stepwise protocol. A total of 1,422 samples were collected over a period of 17 months. The samples were referred for human papillomavirus (HPV) genotyping if they showed cytological evidence of atypical squamous cells of undetermined significance, low- or high-grade squamous intraepithelial lesion. If one or more of HPV types 16, 18, 31, 33, or 45 were present, mRNA was analyzed by the NucliSENS EasyQ HPV assay. The genotypical distribution of high-risk HPV was very heterogeneous; HPV 16, 18, 31, 33, and 45 represented 20.2%, 3.4%, 10.8%, 3.4%, and 3.8% of HPV-positive samples, respectively. Follow-up data were available for 35 patients. Although over the half (51.4%) of follow-up samples showing HPV DNA/mRNA consensus evolved to cervical intraepithelial neoplastic lesions, 25.7% showed no progression to neoplasia despite mRNA positivity. However, the major concern was the group (14.3%) that showed progression to cervical intraepithelial neoplasia despite mRNA negativity: all but one of these cases had a high-risk HPV genotype other than the five included in the NucliSENS EasyQ HPV assay. Markedly, 66.7% of the discordant samples between colposcopy and histology that underestimated the degree of cervical dysplasia were found in this group. Close monitoring of high-risk HPV DNA-positive/mRNA-negative cases remains necessary, which leads to questions about the added value of the evaluated protocol.

Comparing triage algorithms using HPV DNA genotyping, HPV E7 mRNA detection and cytology in high-risk HPV DNA-positive women

“…Previous studies have addressed the value of E6/E7 mRNA NASBA tests for ≥CIN2 detection. These studies, however, used different approaches: primary stratification by, for example, cytology results [13][14][15]17,21,25,[28][29][30][31][32][42][43][44][45] or by HPV16/18 DNA genotyping results [26]. Secondly, these studies addressed NASBA assays targeting five hrHPV types.…”

“…Analysis for E6/E7 transcripts of five hrHPV types (HPV16/18/31/33/45) by commercially available nucleic acid sequence-based amplification (NASBA) tests (PreTect HPV-Proofer, Norchip and NucliSENS EasyQ test, Biomerieux) has been found to identify risk groups amongst hrHPV DNA-positive women with normal cytology [28] and minor cytological abnormalities [34] in need of immediate colposcopy. However, despite its relatively high specificity, the sensitivity of this E6/E7 NASBA test is limited [13,15,16,18,21,[24][25][26]28,34], implying that follow-up of test-negatives remains necessary. Under the assumption that the sensitivity of currently available E6/E7 NASBA tests is restricted by the limited number of targeted hrHPV types, we developed a NASBA assay that allows detection of E7 transcripts of a broader spectrum of hrHPV types (i.e., HPV16/18/31/33/45/52/58).…”