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目的了解贵州吸毒人群丙型肝炎病毒(HCV)感染者的基因亚型分布,为该省吸毒人群HCV临床治疗和估计HCV造成疾病负担提供参考信息。方法抽取贵州省部分美沙酮门诊和强制戒毒所吸毒人员血浆样本,经过初筛和复检,筛选HCV抗体阳性样本;提取HCV核酸后,用巢式RT-PCR扩增5′-UTR区段进行HCV核酸的定性检测(nucleic acids technology,NAT),以筛选HCV核酸阳性样本;再用巢式RT-PCR扩增NS5B区段,对凝胶电泳条带阳性扩增产物进行测序,将清理后的序列与HCVDatabase提供的NS5B区段参考序列比对,使用MEGA 4.0做系统进化树,进行基因分型。结果 309份抗体阳性样本中核酸阳性258份(83.5%),成功分型246份,10份测序区扩增失败,2份测序失败。246份中1a6份,1b25份,3a62份,3b79份,6a69份,6n5份。吸毒人群HCV感染者的基因亚型仅在不同性别和吸毒方式间的分布上差异有统计学意义。NAT(-)与NAT(+)样本HCV抗体酶联免疫吸附法检测S/CO比值差异无统计学意义。结论贵州吸毒人群HCV感染者的基因型主要为3型和6型、少数为1型,未发现2型、4型和5型;性别和吸毒方式影响吸毒人群HCV感染者基因亚型的分布。
Objective To understand the distribution of genotypes of hepatitis C virus (HCV) infected people in Guizhou and to provide reference information for HCV clinical treatment and HCV burden estimation in this province. Methods The plasma samples of methadone clinics and compulsory drug addicts in Guizhou Province were collected and screened for HCV antibody positive samples after primary screening and retesting. After extracting HCV nucleic acids, the 5’-UTR region was amplified by nested RT-PCR for HCV Nucleic acid technology (NAT) was used to screen for positive samples of HCV nucleic acids. The NS5B region was amplified by nested RT-PCR, and the products amplified by gel electrophoresis were sequenced. The cleaned-up sequence Compared with the NS5B segment reference sequence provided by HCVDatabase, the phylogenetic tree was constructed using MEGA 4.0 for genotyping. Results A total of 258 positive samples (83.5%) were obtained from 309 antibody positive samples and 246 were successfully typed. Amplification of 10 sequencing regions failed and 2 of them failed to be sequenced. 246 parts of 1a6 parts, 1b25 parts, 3a62 parts, 3b79 parts, 6a69 parts, 6n5 parts. The genotypes of HCV-infected persons in drug addicts differ only in the distribution between genders and drug users. There was no significant difference in the S / CO ratio between NAT (-) and NAT (+) samples detected by HCV antibody enzyme-linked immunosorbent assay. Conclusion The genotypes of HCV-infected persons in drug users in Guizhou Province are mainly type 3 and type 6, few are type 1, no type 2, type 4 and type 5 are found. The genotypes of genotypes of HCV-infected persons in drug users are affected by gender and drug abuse patterns.