Compared to high-risk human papillomavirus (HPV) DNA detection, detection of E6/E7 transcripts from high-risk HPV genotypes (1) has been proposed as a more specific tool for cervical cancer triage (2). Moreover, highly sensitive transcription-mediated amplification (TMA)-based detection of functional RNA transcripts can theoretically ascribe precursor oncogenicity to HPV-specific nucleic acid. These molecular profiles may precede cellular changes observed in routine cervical cytology. Early assessments of TMA-based detection of E6/E7 transcripts have largely been performed with a digene HC2 high-risk DNA test (HC2; Qiagen, Gaithersburg, MD) predicate device. Reports have described equivalent sensitivity between the two detection modalities, yet improved specificity of E6/E7 detection assays (3-6). In this report, the utility of TMA-based APTIMA HPV (APTIMA; Hologic/Gen-Probe, Incorporated, San Diego, CA) is compared to Cervista HPV HR (Cervista; Hologic, Madison, WI) in the triage of women with underlying cervical intraepithelial neoplasia grade 2ϩ (CIN2ϩ) lesions. Analogous to comparisons with HC2 (3-6), improved specificity was noted with APTIMA, yet our data characterize a phenomenon inherent to Cervista that contributes to deficits in specificity.(Results of this work were previously presented, in part, at the 112th General Meeting of the American Society for Microbiology, San Francisco, CA, 16 to 19 June 2012.)In a study approved by the Wheaton Franciscan Healthcare Institutional Review Board, residual fluid from 4,056 liquid-based cytology specimens (ThinPrep; Hologic, Marlborough, MA) was forwarded for tandem molecular analysis. Inclusion criteria consisted of the following: age of woman of Ն20 years, processing of ThinPrep specimen within 21 days of collection, and presence of sufficient residual fluid for both molecular assays. Glacial acetic acid (GAA)-treated specimens were excluded from analysis, as previous data (7) describe a deleterious effect of GAA on Cervista performance.Matched cytological diagnoses were classified as high-grade squamous intraepithelial lesions (HSIL), low-grade squamous intraepithelial lesions (LSIL), atypical squamous cells of undetermined significance (ASC-US), atypical squamous cells but cannot exclude HSIL (ASC-H), or negative for intraepithelial lesions and malignancy (NILM) (8). When an endocervical biopsy was indicated, histological results were delineated into categories of CIN2ϩ or results of lesser significance.Two-milliliter ThinPrep aliquots were subjected to Cervistabased detection of high-risk HPV types 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 66, and 68 (9). Relative genomic DNA content (gDNA) was assessed by a human histone gene-specific oligonucleotide. DNA extracts were assayed in triplicate; individual luminescence-labeled oligonucleotides were distributed throughout three oligonucleotide mixtures. HPV-specific luminescence from one or two oligonucleotide mixtures was required to equal or exceed a 1.525 ratio to background for a positive result. HPV...
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