反向斑点杂交法快速检测HBV基因型反应条件的优化和建立  

Development of a Reverse Dot Blot System for Rapid Detection of HBV Genotypes

作　　者：赵丽[1] 张文露[1] 胡源[1] 刘彦辰[1] 赖国旗[1] 杨凤[1] 黄爱龙[1] 

机构地区：[1]重庆医科大学感染性疾病分子生物学教育部重点实验室,重庆400016

出　　处：《中国生物工程杂志》2009年第12期85-89,共5页China Biotechnology

基　　金：国家"863"计划资助项目(2007AA02Z400)

摘　　要：利用反向斑点杂交技术,设计出特异性基因分型探针,并将探针固定在带正电荷的尼龙膜上,与PCR扩增带有地高辛标记的临床血清样本进行杂交。通过优化杂交反应条件,建立起简单、快速、特异地检测HBV基因型的方法。利用该方法对重庆地区临床样本进行分型检测,并与直接测序结果比较。结果表明新建的HBV基因分型方法可对拷贝数在103以上的血清样本准确分型,特异性达到96.67%。重庆地区感染HBV主要以B型为主。To develop a reverse dot blot assay for rapid detection of HBV genotypes. Specific oligonucleotides probes were desighed and immobilized on nylon membranes. The DNA sample to be tested was PCR-amplified with DIG labeling primers and then hybridized with the immobilized probes. This procedure for detecting HBV genotypes was simple,rapid and specificity. 30 specimens in Chongqing area were collected and detected by this method, and results were evaluated using direct sequencing. Results showed that： This new method was applicable to precise detection HBV genotypes for specimen with copies up to 10^3, and the HBV genotyping results showed that genotype B was the predominant genotype in Chongqing area.

关 键 词：反向斑点杂交 HBV 基因型 临床样本 

分 类 号：Q819[生物学—生物工程]