Laboratory of Molecular Genetics and Biotechnology (GMBio), Department of Biology, Center for Biological and Health Sciences, Federal University of Sergipe, São Cristóvão, Brazil.
J Med Virol. 2021 Nov;93(11):6347-6354. doi: 10.1002/jmv.27118. Epub 2021 Jun 7.
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.
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.
The procedures were based on different primer design strategies, using MY09/MY11, EntroA, and type specific multiplex PCR primers.
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.
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.
人乳头瘤病毒（HPV）是宫颈癌的主要病因。聚合酶链反应（PCR）为基础的技术与 HPV 检测和基因分型的准确结果相关联，能够以低水平识别病毒 DNA。然而，不同的引物设计会影响其敏感性和特异性，具体取决于评估的 HPV 类型。
本研究旨在比较评价三种不同的基于 PCR 的策略在检测和分型宫颈样本中的 HPV 的有效性。
该程序基于不同的引物设计策略，使用 MY09/MY11、EntroA 和型特异性多重 PCR 引物。
在 411 例宫颈刮片样本中，分别有 45（10.9%）、50（12.2%）和 117（28.5%）例样本的 MY09/MY11、EntroA 和多重 PCR 检测为阳性。对于 MY09/MY11 阳性样本，36 例样本 EntroA 检测为阴性，23 例多重 PCR 检测为阴性。对于 EntroA 阳性样本，40 例样本 MY09/MY11 检测为阴性，26 例多重 PCR 检测为阴性。对于多重 PCR 阳性样本，96 例样本 MY09/MY11 检测为阴性，94 例样本 EntroA 检测为阴性。MY09/MY11 鉴定出 12 种不同的 HPV 类型，EntroA 检测出 8 种类型，多重 PCR 检测出 11 种 HPV 类型。EntroA 引物能够检测到更多的 HPV 样本，而多重 PCR 尽管针对的 HPV 类型有限，但敏感性高于其他方法。
三种方法各有优缺点，本研究强调需要使用一种以上的分子策略进行 HPV 检测和基因分型，并开发新的方法来克服现有检测方法的局限性。
