摘要
目的针对乙肝病毒(HBV)不同亚型,建立环介导等温扩增(loop-mediated isothermal amplification,LAMP)方法,实现对HBV的快速检测及基因分型。方法分别设计HBV B、C、D三亚型基因组特征序列的LAMP引物,优化反应条件,建立特异灵敏的HBV亚型LAMP检测方法,并与Real-time PCR方法进行临床样本检测结果比对。结果 LAMP法在数十分钟内有效地检测出HBV的B、C、D三亚型;其中,HBV-B亚型的检出限为17拷贝;C亚型的检出限为25拷贝;D亚型的检出限为10拷贝。与Real-time PCR法相比,灵敏度提高1 000倍左右。结论 LAMP法可实现HBV的快速检测及分型,具有高特异度和高灵敏度,可用于临床检测诊断。
Objective A loop-mediated isothermal amplification（LAMP）method had been developed to detect different subtypes of hepatitis B virus（HBV）rapidly.Methods LAMP primers were designed according to the specific DNA sequences of HBV B,C and D genotypes,respectively.LAMP reactions were carried out with optimal conditions.The specificity of LAMP primer sets was validated among HBV subtypes of B,C and D.The specificity and sensitivity of LAMP were compared with those of real-time PCR by assaying clinical samples of HBV.Results LAMP method could detect HBV B,C and D subtype effectively in tens of minutes and the detection limits of HBV-B subtype were 17 copies,HBV-C subtype 25 copies and HBV-D subtype 10 copies.Compared with real-time PCR,the sensitivity of LAMP was enhanced by about 1 000 times.Conclusion LAMP could detect different HBV genotypes rapidly with high sensitivity and specificity.
出处
《华中科技大学学报（医学版）》
CAS
CSCD
北大核心
2012年第1期49-53,共5页
Acta Medicinae Universitatis Scientiae et Technologiae Huazhong
基金
武汉市科技局对外科技合作与交流计划项目(No.201070934341)
科技攻关计划项目
武汉工业学院研究生创新基金重点项目(No.09cx002)
关键词
乙型肝炎病毒
亚型
环介导等温扩增法
hepatitis B virus
subtype
loop-mediated isothermal amplification
