Centro de Desenvolvimento Científico e Tecnológico, Fundação Estadual de Produção e Pesquisa em Saúde (CDCT/FEPPS), Porto Alegre, RS, Brazil.
J Virol Methods. 2011 Oct;177(1):38-43. doi: 10.1016/j.jviromet.2011.06.010. Epub 2011 Jul 22.
Persistent infection with high-risk human papillomavirus (HR-HPV) has been associated with cervical cancer. Developing assays for the identification of these viral types is of great importance for monitoring patients and controlling strategies. The development of the MCHA (microplate colorimetric hybridization assay), a PCR-based method for identifying six of the most common HR-HPV types (HPV 16, 18, 31, 33, 39 and 45) is described. The MCHA combines the amplification with the GP5+/GP6+ consensus primers followed by PCR reverse hybridization with specific probes and detection through a colorimetric assay. The performance of the MCHA was evaluated using 108 DNA samples typed previously by the PapilloCheck(®). The agreement between both methods was 69.4% for HPV 16; 79.1% for HPV 45; 82.4% for HPV 18; 93.6% for HPV 31; 87.9% for HPV 33, and 17.6% for HPV 39. The assay had higher sensitivity than the Papillocheck(®), particularly for identifying HPV 16 and 18. The MCHA seemed to be sensitive and specific for the identification of the most prevalent HPV types in invasive cervical cancer, HPV 16, 18, 45, 33 and 31. It requires low-cost reagents and common laboratory apparatus.
高危型人乳头瘤病毒（HR-HPV）的持续感染与宫颈癌有关。开发用于鉴定这些病毒类型的检测方法对于监测患者和控制策略非常重要。本文描述了一种基于 PCR 的 MCHA（微孔板比色杂交检测法）的开发，该方法可鉴定六种最常见的 HR-HPV 类型（HPV 16、18、31、33、39 和 45）。MCHA 将扩增与 GP5+/GP6+ 通用引物相结合，然后进行 PCR 反向杂交与特异性探针，并通过比色检测进行检测。使用 PapilloCheck(®)先前分型的 108 个 DNA 样本评估了 MCHA 的性能。两种方法之间的一致性为：HPV 16 为 69.4%；HPV 45 为 79.1%；HPV 18 为 82.4%；HPV 31 为 93.6%；HPV 33 为 87.9%；HPV 39 为 17.6%。该检测方法的敏感性高于 Papillocheck(®)，特别是对 HPV 16 和 18 的鉴定。MCHA 似乎对鉴定侵袭性宫颈癌中最常见的 HPV 类型 HPV 16、18、45、33 和 31 具有敏感性和特异性。它需要低成本的试剂和常见的实验室设备。
