摘要
为研究梯型熔解温度等温扩增(LMTIA)技术检测人乳头瘤病毒(HPV)核酸的可行性,分析高危型HPV16、HPV18、HPV35、HPV39、HPV45、HPV51、HPV52、HPV56和HPV58的基因组,选择熔解温度曲线呈梯型的特异性序列,合成特异性序列并克隆到pUC57载体上作为DNA模板,通过设计LMTIA引物、优化反应温度来测定灵敏度.结果表明,所建立的9种HPV高危型病毒LMTIA检测方法在最适温度下稳定性强,对pUC57质粒模板DNA的检测灵敏度为0.1~1 fg/reaction.LMTIA方法稳定性强、灵敏度高,在检测HPV核酸方面具有广阔的应用前景.
To study the feasibility of detecting the human papillomavirus(HPV)with ladder-type melting temperature isothermal amplification(LMTIA)technology,the genomes of high-risk HPV16,HPV18,HPV35,HPV39,HPV45,HPV51,HPV52,HPV56 and HPV58 were analyzed and the specific sequences with ladder-type melting temperature curves were selected.These sequnces were sythensized and coloned into pUC57 vectors as DNA templates.Through the LMTIA primers were designed,the reaction temperature was optimzed,and the detection limits were determined.The established 9 LMTIA methods for detection of high-risk HPV strains were of high roboustness at optimized temperature,and the detection limits for pUC57 vector templates were 0.1-1 fg/reaction.The LMTIA methods are of high repeatability and sensitivity,which have great application potiential in detection of HPV.
作者
孔爱华
艾旭光
王德国
KONG Aihua;AI Xuguang;WANG Deguo(People’s Hospital of Dingtao Distric,Heze 274100,China;Key Laboratory of Biomarker Based Rapid-Detection Technology for Food Safety of Henan Province/Food and Pharmacy College,Xuchang University,Xuchang 461000,China)
出处
《许昌学院学报》
CAS
2022年第2期72-75,共4页
Journal of Xuchang University
基金
国家重点研发计划项目(2016YFD0500704)
国家自然科学基金项目(31701689,32172300)。
关键词
梯型熔解温度等温扩增
人乳头瘤病毒
核酸检测
靶序列
引物设计
Ladder-type Melting Temperature Isothermal amplification
human papillomavirus
nucleic acids detection
target sequence
primer design
作者简介
孔爱华(1975-),女,山东定陶人,副主任医师,研究方向:呼吸内科方向;通讯作者:王德国(1975-),男,山东菏泽人,教授,博士,研究方向:基因快速检测.
