吉林大学学报(理学版)

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人乳头瘤病毒16和52型双荧光等温多自配引发扩增的

崔玉伟1, 牟颖2, 马莉1, 丁雄2, 方宗宇1, 王焰1, 吴青青1   

  1. 1. 贵州医科大学附属医院 中心实验室, 贵阳 550004;2. 浙江大学 工业控制技术国家重点实验室, 智能系统与控制研究所, 分析仪器研究中心, 杭州 310058
  • 收稿日期:2017-12-18 出版日期:2018-07-26 发布日期:2018-07-31
  • 通讯作者: 吴青青 E-mail:wuqingqing@gmc.edu.cn

Detection Method of Human Papillomavirus HPV16 and HPV52by Dual Fluorescence IsothermalMultipleSelfMatchingInitiated Amplif

CUI Yuwei1, MU Ying2, MA Li1, DING Xiong2, FANG Zongyu1, WANG Yan1, WU Qingqing1   

  1. 1. Central Laboratory of Affiliated Hospital of Guizhou Medical University, Guiyang 550004, China; 2. Research Center for Analytical Instrumentation, Institute of CyberSystems and Control, State Key Laboratory of Industrial Control Technology, Zhejiang University, Hangzhou 310058, China
  • Received:2017-12-18 Online:2018-07-26 Published:2018-07-31
  • Contact: WU Qingqing E-mail:wuqingqing@gmc.edu.cn

摘要: 应用等温多自配引发扩增(IMSA)技术, 分别针对人乳头瘤病毒(HPV)16型的E7和52型的E6基因序列设计6条特异性引物, 并在检测体系中加入羟基萘酚蓝(HNB)和SYBR GreenⅠ的混合双荧光指示剂, 建立快速检测人乳头瘤病毒的双荧光IMSA方法. 结果表明: 340 μmol/L HNB与1∶10 000 SYBR GreenⅠ混合构建的双荧光指示剂在IMSA反应体系中具有明确的指示效果, 455 nm蓝光激发下阳性反应管双荧光显色为黄绿色, 阴性反应管双荧光显色为橘红色; 该方法对HPV16和HPV52型检测限分别达60,600拷贝/μL, 可特异性检出样品中HPV16和HPV52, 与临床检测结果比对无差异.

关键词: 人乳头瘤病毒16型, 人乳头瘤病毒52型, 等温多自配引发扩增, 双荧光

Abstract: The isothermal multipleselfmatchinginitiated amplification (IMSA) technology was used to detect human papillomavirus (HPV) and six specific primers were designed for HPV16E7 or HPV52E6 gene sequences. Meanwhile, dual fluorescence indicator (hydroxy naphthol blue (HNB) and SYBR Green Ⅰ) was added to the detection system. We established dual fluorescence IMSA method for rapid detection of human papillomavirus. The results show that the dual fluorescence indicator constructed by 340 μmol/L HNB and 1∶10 000 SYBR Green Ⅰ has obvious indication effect in the IMSA reaction system. The coloration of positive reaction tube is yellowgreen under the 455 nm blue excitation,  the coloration of negative reaction tube is orangered, and the detection limits of HPV16 and HPV52 are 60,600 copies/μL, respectively, HPV16 and HPV52 can be specifically detected in the samples, and there is no difference in comparison with clinical detection results.

Key words: dual fluorescence, human papillomavirus 52 (HPV52), human papillomavirus 16 (HPV16), isothermal multipleselfmatchinginitiated amplification (IMSA)

中图分类号: 

  • Q31