套式PCR检测抗-HBs阳性者血清HBVDNA及序列分析  

Detection of HBVDNA in serum from patients with anti - HBs by nested - PCR and analysis of nudeotide.

作　　者：黄利华[1] 蒋跃明[1] 裴浩[1] 魏建国[1] 曹立森[1] 

机构地区：[1]无锡市传染病医院,江苏无锡214007

出　　处：《临床肝胆病杂志》2003年第2期102-103,共2页Journal of Clinical Hepatology

基　　金：无锡市2000年科学技术发展计划项目(批号WZ2000192)

摘　　要：研究套式PCR在检测抗-HBs阳性不同模式者血清HBVDNA中的意义,从基因角度分析产生的原因。对64例标本采用套式PCR检测HBVDNA并作序列分析,荧光定量法检测HBVDNA含量。抗-HBs阳性不同模式者常规PCR法检测HBVDNA阳性率29.7％,套式PCR检测阳性率84.4％;套式PCR阳性组肝功能损害严重、HBVD-NA与HBsAg值显著高于套式PCR阴性组;套式PCR性阴/常规PCR阴性组48.6％的病例由于S基因变异导致HB-sAg肽氨基酸置换产生的,抗-HBs不能中和病毒而与HBVDNA共存。因此对那些抗-HBs阳性、肝功能反复异常者有必要进行套式PCR检测,以提高HBV感染的诊断率。To discuss the significance of nested polymerase chain reaction (nested - PCR) in patients with hepatitis B surface antibody (anti - HBs)and to analysis cause of it with gene. Quantitative analysis of HBVDNA from 64 patients was performed by PCR. Qualitative analysis of HBVDNA was tested by nested - PCR and S gene fragments were directly sequenced. Positive rate of HBVDNA in serum was 29.7% by routine PCR in comparision with 84.4% by nested - PCR in patients with different patterns of anti - HBs position. Liver function was more severe and HBVDNA, HBsAg tilers were significantly higher in nested - PCR positive group than in nested - PCR negative group. Amino acid substitutions of HBsAg were delected in patients in nested - PCR positive while routine PCR negative group, and include the Mowing: THr47L, THr63S, L82A, L89A, C90S, L91A, H101A,THr115S, I126A,C139G,S154A. These findings suggested that mutation located in 'a' determinant cluster and two extremity near to it may result in a conformation change of the S protein, and affect its antigenicities, then cause coexist of HBVDNA and anti - HBs. Therefore, it was necessary to detect serum HBVDNA by nested - PCR for patients with anti - HBs while liver function abnormal in order to improve diagnosis of HBV infection.

关 键 词：套式PCR HBVDNA 抗-HBS 基因变异 

分 类 号：R446.5[医药卫生—诊断学]