流行高危人乳头瘤病毒的快速多重PCR检测

A quick multiplex PCR method for the detection of high risk human papillomavirus

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。

The human papillomavirus （HPV） type 16, 18, 58 are the most prevalent high risk types in Shaanxi Province. The study is intended to establish a simple, inexpensive and time-saving techniques for detecting and typing HPV16, 18, 58. Type specific primers for HPV16, 18, 58 were designed according to the genomic DNA se- quences provided by GeneBank and synthesized, the conditions of PCR were optimized, the plasmids containing HPV 16, 18, 58 genome respectively were used as positive controls, the cervical cancer samples with predeter- mined HPV infection were assayed by the novel multiple PCR technique. The results showed that this method allows detecting and typing HPV16, 18, 58 DNA simultaneously in one reaction based on the length of the PCR products after electrophoresis. The novel method is reliable and can effectively detect multiple HPV infection within the le- sions. Using the single PCR reaction, HPV 16, 18 and 58 were successfully identified from samples known to be positive for these HPV types. This simplified, economic and time-saving multiplex PCR method provides a useful tool for HPV screening.