Cavalcante Gustavo Henrique O, de Araújo Josélio M G, Fernandes José Veríssimo, Lanza Daniel C F
Applied Molecular Biology Lab - LAPLIC, Department of Biochemistry, Federal University of Rio Grande do Norte, Natal, RN, Brazil; Postgraduate Program in Biochemistry, Federal University of Rio Grande do Norte, Natal, RN, Brazil.
Laboratory of Molecular Biology for Infectious Diseases and Cancer - LADIC, Department of Microbiology and Parasitology, Federal University of Rio Grande do Norte, Natal, RN, Brazil.
Diagn Microbiol Infect Dis. 2018 May;91(1):20-26. doi: 10.1016/j.diagmicrobio.2017.12.016. Epub 2017 Dec 24.
HPV infection is considered one of the leading causes of cervical cancer in the world. To date, more than 180 types of HPV have been described and viral typing is critical for defining the prognosis of cancer. In this work, a seminested PCR which allow fast and inexpensively detection and typing of HPV is presented. The system is based on the amplification of a variable length region within the viral gene E1, using three primers that potentially anneal in all HPV genomes. The amplicons produced in the first step can be identified by high resolution electrophoresis or direct sequencing. The seminested step includes nine specific primers which can be used in multiplex or individual reactions to discriminate the main types of HPV by amplicon size differentiation using agarose electrophoresis, reducing the time spent and cost per analysis.
人乳头瘤病毒（HPV）感染被认为是全球宫颈癌的主要病因之一。迄今为止，已发现超过180种HPV类型，病毒分型对于确定癌症预后至关重要。在这项研究中，我们提出了一种半巢式聚合酶链反应（PCR），它能够快速且低成本地检测HPV并进行分型。该系统基于病毒E1基因内可变长度区域的扩增，使用三个可能与所有HPV基因组退火的引物。第一步产生的扩增子可通过高分辨率电泳或直接测序进行鉴定。半巢式步骤包括九个特异性引物，可用于多重或单独反应，通过琼脂糖电泳根据扩增子大小差异来区分主要的HPV类型，从而减少每次分析所需的时间和成本。
