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HPV16,18,58是陕西地区最为流行的三型高危人乳头瘤病毒(human papillomavirus HPV)株,拟建立针对流行高危HPV型别的快速筛查方法。根据Genebank提供的序列,设计并合成HPV16,18,58型特异型引物,优化PCR条件,以含有HPV16,18,58全基因组序列的质粒为阳性对照模板,并以预先确定HPV感染状态的宫颈癌组织标本验证所建立的方法。该方法可以成功地一次扩增HPV16,18,58三型DNA片断,并经琼脂糖凝胶电泳解析。应用预先确证HPV16,18,58多重感染的宫颈癌组织标本为模板,也成功地扩增出相应的HPV基因片断。建立的单次多重PCR方法简单、经济、节约时间,可望用于当地流行高危型HPV16,18,58的群体筛查。
HPV16,18,58 is the most popular type III human papillomavirus strain in Shaanxi Province. It is planned to establish a rapid screening method for the high-risk HPV types. Based on the sequences provided by Genebank, HPV16, 18, and 58 specific primers were designed and synthesized, and the PCR conditions were optimized. The plasmid containing HPV16, 18 and 58 whole genome sequences was used as a positive control template. Cervical cancer with HPV infection status Tissue specimen verification established by the method. This method can successfully amplify HPV16,18,58 type III DNA fragment at a time and analyze by agarose gel electrophoresis. HPV16,18,58 multiple infection of cervical cancer tissue samples confirmed as a template, but also successfully amplified the corresponding HPV gene fragments. The established single multiplex PCR method is simple, economical and time-saving, and is expected to be used for population screening of locally-endemic high-risk HPV16,18,58.