荧光定量PCR法分析乙肝病毒基因型及载量  

Analysis of genotype and quantification of hepatitis B virus by real-time PCR assay

作　　者：刘琦[1] 曾爱中[1] 秦波[1] 赵耀[2] 胡接力[3] 黄爱龙[3] 

机构地区：[1]重庆医科大学附属第一医院感染科,重庆400016 [2]重庆医科大学附属儿童医院儿科研究所,重庆400014 [3]重庆医科大学教育部感染性疾病分子生物学重点实验室病毒性肝炎研究所,重庆400016

出　　处：《重庆医科大学学报》2011年第7期829-832,共4页Journal of Chongqing Medical University

基　　金：国家自然科学基金资助项目(编号:30972587)

摘　　要：目的：了解重庆地区乙型肝炎病毒（Hepatitis B virus,HBV）基因型的分布情况并进行精确定量分析。方法：采用荧光分型定量聚合酶链反应法（Real-time genotyping and quantitative PCR,GQ-PCR）对收集的血清标本进行HBV基因分型并定量。结果：152株血清标本成功分型148株（97.37%）,其中B基因型109例（男88例、女21例）[定量值范围4-12 log10copies/ml、均值和标准差（7.94±1.44）log10copies/ml]、C基因型10例（男6例、女4例）[定量值范围5~10 log10copies/ml、均值和标准差（8.12±1.64）log10copies/ml]、基因B＋C混合型29例（男24例、女5例）[定量值范围4~10 log10copies/ml、均值和标准差（7.45±1.59）log10copies/ml]。检出率分别为73.65%（男74.58%、女70.00%）、6.76%（男5.08%、女13.30%）、19.59%（男20.34%、女16.70%）。结论：重庆地区HBV优势基因型以B型和B＋C混合型为主,与前几年相比较,C型有下降趋势。基因型的分布与性别无关（P〉0.05）。HBV载量在B、C型及B＋C混合型三者之间无统计学差异（P〉0.05）。荧光分型定量PCR（GQ-PCR）法灵敏度和特异性高,能够同时准确的对临床标本进行HBV基因分型及定量检测,并有望替代传统的基因分型方法。Objective： To make precise quantitation and distribution analysis of hepatitis B virus（HBV） genotypes in Chongqing.Methods： Real-time genotyping and quantitative PCR（GQ-PCR） was used to genotype and quantitate the HBV strains.Results： A total of 148（97.37%）out of the 152 strains were successfully genotyped.Among them,109 patients（88 males and 21 females） had genotype B with quantity ranging from 4~12 log10copies/ml,and mean and standard deviation of（7.94±1.44） log10copies/ml,10 patients（6 males and 4 females） had genotype C with quantity ranging from 5~10 log10copies/ml,and mean and standard deviation of（8.12±1.64） log10copies/ml,29 patients（24 males and 5 females） had mixtures of genotype B and C with quantity ranging from 4~10 log10copies/ml,and mean and standard deviation of（7.45±1.59） log10copies/ml.The detecting rates of genotype B,genotype C,and genotype B and C were 73.65%（74.58% in males,70.00% in females）,6.76%（5.08% in males,13.30% in females）,and 19.59%（20.34% in males,16.70% in females）,respectively.Conclusion： Genotype B is the most dominant type for HBV strains,followed by mixed genotype B and C infection.Genotype C decreased compared with the previous years.There is no relationship between the gender and HBV genotypes.No significant difference was found between HBV genotype and HBV DNA level.GQ-PCR has a higher sensitivity and specificity for precise simultaneous genotyping and quantifing HBV clinical samples and can replace the traditional genotyping assays.

关 键 词：乙型肝炎病毒 HBV基因型 荧光定量PCR 

分 类 号：R512.62[医药卫生—内科学]