Simultaneous detection of hepatitis B virus genotypes and mutations associated w

StephenW

Braz J Infect Dis. 2011 Dec;15(6):560-6.
Simultaneous detection of hepatitis B virus genotypes and mutations
associated with resistance to lamivudine, adefovir, and telbivudine by the
polymerase chain reaction-ligase detection reaction. Source:
http://www.ncbi.nlm.nih.gov/pubmed/22218515 Wang YZ, Xiao JH, Liu LG, Ye
CY, Shen HY, Xu TM, Zhu KZ. SourceInstitute for the Study of Liver
Diseases, The Third People's Hospital of Changzhou, China.

Abstract
OBJECTIVES: Detection of mutations associated to nucleos(t)ide analogs and
hepatitis B virus (HBV) genotyping are essential for monitoring treatment
of HBV infection. We developed a multiplex polymerase chain reaction-ligase
detection reaction (PCR-LDR) assay for the rapid detection of HBV
genotypes and mutations associated with lamivudine, adefovir, and
telbivudine resistance in HBV-infected patients. METHODS: HBV templates
were amplified by PCR, followed by LDR and electrophoresis on a sequencer.
The assay was evaluated using plasmids that contained wild-type or mutant
HBV sequences and 216 clinical samples. RESULTS: The PCR-LDR assay and
sequencing gave comparable results for 158 of the 216 samples (73.1%) with
respect to mutation detection and genotyping. Complete agreement between
the two methods was observed for all the samples (100%) at codon 180 and
codon 204. Concordant results were observed for 99.4% of the 158 samples at
codon 181 and 98.7% at codon 236. The genotyping results were completely
concordant between the PCR-LDR assay and sequencing. The PCR-LDR assay
could detect a proportion of 1% mutant plasmid in a background of wild-type
plasmid. CONCLUSION: The PCR-LDR assay is sensitive and specific for
detection of HBV genotypes and drug resistance mutations, and could be
helpful for decision making in the treatment of HBV infection.

StephenW

Braz传染病杂志。 2011十二月15（6）:560 -6。
同时检测乙型肝炎病毒基因型和突变
与耐拉米夫定，阿德福韦，替比夫定由
聚合酶链反应 - 连接酶检测反应。来源：
http://www.ncbi.nlm.nih.gov/pubmed/22218515王YZ LG，叶，刘，肖JH
CY，沉煜，徐TM，朱KZ。 SourceInstitute为肝脏研究
疾病，中国常州市第三人民医院。

摘要
目标：核苷（酸）IDE类似物相关的基因突变检测和
B型肝炎病毒（HBV）基因分型是必不可少的监测治疗
乙肝病毒感染。我们开发了一种多重聚合酶链反应 - 连接酶
检测的反应（PCR - LDR）检测为乙肝病毒的快速检测
基因型和突变与拉米夫定，阿德福韦，并
替比夫定耐药性HBV感染患者。方法：乙肝病毒模板
LDR和音序电泳PCR扩增。
使用含有野生型或突变的质粒检测评价
HBV序列和216个临床样本。结果：PCR - LDR检测和
测序了比较的结果（73.1％）的216个样本158
尊重突变检测和基因分型。之间的完整协议
两种方法观察180位密码子和所有样品的（100％）
密码子204。一致的结果，观察的158个样本的99.4％
密码子181密码子236和98.7％。基因分型结果完全
之间的PCR- LDR检测和测序一致。 PCR - LDR检测
可以检测的1％的突变质粒比例，在野生型背景
质粒。结论：PCR - LDR检测的敏感性和特异性
HBV基因型和耐药突变检测，并可以
在乙肝病毒感染的治疗决策有帮助的。