Comparative analysis of different PCR‐based strategies for HPV detection and genotyping from cervical samples

Abstract Background Human papillomavirus (HPV) is the main cause of cervical cancer. Polymerase chain reaction (PCR)‐based techniques are associated with accurate results with respect to HPV detection and genotyping, being able to identify viral DNA at low levels. However, differences in primer design influence their sensibility and specificity, depending on the HPV type assessed. Objective The aim of the study was to comparatively evaluate the effectiveness of three different PCR‐based strategies for HPV detection and genotyping from cervical samples. Study Design The procedures were based on different primer design strategies, using MY09/MY11, EntroA, and type specific multiplex PCR primers. Results Out of 411 samples of cervical scrapings, 45 (10.9%), 50 (12.2%), and 117 (28.5%) were positive for MY09/MY11, EntroA, and multiplex PCR, respectively. For MY09/MY11 positive samples, 36 were negative for EntroA and 23 for multiplex PCR. For EntroA positive samples, 40 were negative for MY09/MY11 and 26 for multiplex PCR. For multiplex PCR positive samples, 96 were negative for MY09/MY11 and 94 for EntroA. MY09/MY11 identified 12 different HPV types, EntroA detected eight types and multiplex PCR detected 11 HPV types. EntroA primers were able to detect HPV in more samples than MY09/MY11, while multiplex PCR, despite the limited targeted HPV types, presented higher sensibility than the other methods. Conclusions The three methods presented different advantages and disadvantages, and the present study reinforces the need to use more than one molecular strategy for HPV detection and genotyping, and the development of novel methods which could overcome the limitations of the existing tests.