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目的探讨江苏地区献血者感染的HCV基因型及亚型。方法收集2013年来自江苏省血液中心ELISA双试剂检测抗-HCV阳性献血者血清标本,使用HCV核酸定量检测试剂盒检测HCV RNA载量;同时使用RNA提取试剂提取HCV RNA,反转录PCR法扩增HCV Core区基因片段,并对扩增产物进行测序,利用进化树分析基因型和亚型。结果 2013年抗-HCV阳性标本139例(0.20%),其中64例抗-HCV阳性血清标本经荧光定量检测,RNA阴性30份,阳性34份。经巢式PCR扩增能够分型的标本24份,包括1a(71.7%)、1b(7.5%)、2a(7.5%)和3b(1.9%)3种基因型和4种亚型。年龄偏大(>35岁)以及男性的HCV RNA阳性标本的病毒滴度与抗-HCV水平(S/CO值)都高于RNA阴性标本,并且存在性别差异,但年龄间无差异。结论江苏无偿献血者感染的HCV基因型以1型为主,其中1b为优势基因亚型。病毒血症献血者的HCV抗体水平显著升高,HCV RNA在自然感染进程中的变化有待通过进一步随访进行研究。
Objective To investigate the genotypes and subtypes of HCV in blood donors in Jiangsu Province. Methods Serum samples of anti-HCV positive blood donors were collected from Jiangsu Blood Center ELISA dual-reagent in 2013. HCV RNA was detected by HCV nucleic acid quantitative detection kit. Meanwhile, HCV RNA was extracted by RNA extraction reagent and reverse transcription PCR The HCV core region gene fragment was amplified, and the amplification product was sequenced. The genotypes and subtypes were analyzed by using the evolutionary tree. Results In 2013, 139 anti-HCV positive samples (0.20%) were detected. Among them, 64 anti-HCV positive serum samples were detected by fluorescence quantitative analysis, with 30 RNA negative and 34 positive. Twenty-four specimens were genotyped by nested PCR, including 3 genotypes and 4 subtypes of 1a (71.7%), 1b (7.5%), 2a (7.5%) and 3b (1.9%). The virus titer and anti-HCV level (S / CO value) of older HCV genotypes (> 35 years old) and male HCV RNA positive samples were higher than those of RNA negative samples, and there were gender differences but no difference between the ages. Conclusion The genotypes of HCV infected by blood donors in Jiangsu are mainly type 1, among which 1b is the predominant genotype. HCV antibody levels are significantly increased in viremic blood donors, and changes in HCV RNA during natural infection remain to be investigated through further follow-up.