Human papillomavirus (HPV) DNA genotyping is an essential test to establish efficacy in HPV vaccine clinical trials and HPV prevalence in natural history studies. A number of HPV DNA genotyping methods have been cited in the literature, but the comparability of the outcomes from the different methods has not been well characterized. Clinically, cytology is used to establish possible HPV infection. We evaluated the sensitivity and specificity of HPV multiplex PCR assays compared to those of the testing scheme of the Hybrid Capture II (HCII) assay followed by an HPV PCR/line hybridization assay (HCII-LiPA v2). SurePath residual samples were split into two aliquots. One aliquot was subjected to HCII testing followed by DNA extraction and LiPA v2 genotyping. The second aliquot was shipped to a second laboratory, where DNA was extracted and HPV multiplex PCR testing was performed. Comparisons were evaluated for 15 HPV types common in both assays. Cervical cancer is the second most common cause of cancer death among women globally. An estimated 510,000 new cases will be diagnosed around the world this year, and 288,000 women will die from the disease (15). In developing countries, cervical cancer can be the most common cancer among women. There is a strong causal relationship between infection with human papillomavirus (HPV) and cervical cancer, as the prevalence of the virus in cervical cancers has been estimated to be as high as 99.7% (14).Reliable, sensitive, and accurate detection of specific HPV types is critical for the study of cervical cancer causation as well as the support of vaccine clinical trials. Several PCR-based methodologies have been developed to amplify and detect HPV DNA (1,4,7,8,9,12, 13). Many of these methodologies exploit the high homology found within specific open reading frames (ORFs) across different HPV types through the use of consensus or degenerate primer pairs that are capable of PCR amplifying numerous HPV types present in a sample. The resultant amplicons are detected and genotyped by utilizing HPV type-specific probes and colorimetric precipitation. While a greater number of HPV types can be detected by these methodologies, additional experimentation beyond PCR cycling is required to identify specific HPV types. Concerns arise over the increased contamination risk with postamplification experimentation and equivalent optimization between HPV types using single-amplification and single-hybridization conditions.For efficacy determinations of the recently FDA-approved quadrivalent HPV (types 6, 11, 16, 18) L1 VLP vaccine (Gardasil; Merck & Co., Inc.) (2, 3), highly sensitive and specific HPV multiplex PCR assays were developed (5, 10, 11) and used, which simultaneously detect the L1 and/or E6 and E7 ORFs of a specific HPV type using the ABI Prism 7700 sequence detection system instrument (Applied Biosystems, Foster City, CA). In this system, DNA is purified from multiple anogenital swab samples per patient by utilizing spin column chemistry under very stringent conditions. All samples ...
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