PCR-反向斑点杂交法检测HBV YMDD模序混合感染  

PCR-Flow Through RDB is A Rapid and Effective Assay for HBV YMDD Motif Mixed Infection

作　　者：袁耀钦 潘小划 何东华[2] 欧志英[3] 

机构地区：[1]广东省东莞市茶山医院,东莞523382 [2]中山大学达安基因诊断中心,广州510080 [3]广州市儿童医院,广州510120

出　　处：《热带医学杂志》2007年第11期1064-1067,共4页Journal of Tropical Medicine

基　　金：国家863科技攻关计划(No.2002AA215018)。

摘　　要：目的为HBV感染临床个体化治疗、指导临床用药寻找一种能更好地检测HBV YMDD模序混合感染的方法。方法28份已知乙肝病人血清,其中10份为HBV YMDD模序感染,5份为HBV YIDD模序感染,5份为HBV YVDD模序感染,2份为HBV YMDD和YIDD模序混合感染,2份为HBV YMDD和YVDD模序混合感染,2份为HBV YIDD和YVDD模序混合感染,2份为HBV YMDD、YIDD和YVDD模序混合感染,都用PCR产物直接测序和PCR-流过式反向斑点杂交(Flowthrough-RDB)方法检测。结果HBV YMDD模序单一感染标本用两种方法检测准确性都达到100%,对混合感染标本PCR-流过式反向斑点杂交方法检测准确性也可达100%,然而用PCR产物直接测序方法只能检测其中一种或两种型别。结论PCR-流过式反向斑点杂交方法是一种敏感、特异和快速的检测HBV YMDD野生型及其突变型模序单一感染和混合感染的有效方法,比PCR产物直接测序更优越,更有利于实现临床个体化治疗和指导临床用药。Objective To develop a better method for the identification of HBV YMDD motif mixed infection, and provide individualized treatment and clinical medication guideline. Methods Twenty-eight archival known sera from chronic HBV carrier, of which 10 were HBV YMDD motif, 5 were HBV YIDD motif, 5 were HBV YVDD motif, 2 were mixed infection with HBV YMDD and YIDD motif, 2 were mixed infection with HBV YMDD and YVDD motif, 2 were mixed infection with HBV YIDD and YVDD motif and 2 were mixed infection with HBV YMDD, YIDD and YVDD motif, were detected by PCR based DNA direct sequencing and PCR-flow through reverse dot blot （RDB）hybridization assay. Results Both PCR-flow through RDB hybridization assay and PCR based DNA direct sequencing were 100% accurate for HBV mono-infection; PCR-flow through RDB hybridization assay is 100% accurate but the PCR based DNA direct sequencing is insufficient to detect all the genotypes of the mixed infection. Conclusion PCR-flow through RDB hybridization is a better detection method for HBV YMDD motif mixed infection than PCR based DNA direct sequencing. It is a better method for genotyping HBV and provides guideline for individualized treatment and clinical medication.

关 键 词：HBV 拉米夫定 PCR-流过式反向斑点杂交 测序 

分 类 号：R512.62[医药卫生—内科学]