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系统进化树在维持性血液透析患者丙型肝炎病毒感染的基因型及同源性分析中的应用
Polygenetic Tree Analysis of hepatitis C virus genotype in hemodialysis patients
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DOI:
作者:
陈嘉
Jia Chen,Shaodong Luan,Yongcheng He,Ding Xiaoqiang
作者单位:
深圳市第二人民医院肾内科 复旦大学中山医院肾内科
The Second People’s Hospital of Shenzhen The Zhongshan Hospital of Fudan University
关键词:
丙型肝炎病毒;基因型;血液透析;系统进化树
Hepatitis C virus; Genotype; Hemodialysis; Phylogenetic tree
摘要:
目的 通过对深圳市第二人民医院血液透析中心抗-丙型肝炎病毒(抗-HCV)及HCV-RNA双阳性的维持性血液透析(MHD)患者血清进行系统进化树分析,明确HCV阳性患者基因型及亚型,并对HCV阳性患者的同源性进行分析,为透析中心HCV感染的防治提供依据。方法 荧光定量聚合酶链反应(PCR)法测定抗-HCV阳性患者病毒载量,对HCV-RNA≥1000U/mL的患者,通过巢氏反转录(RT)-PCR对5’UTR及NS5B区进行扩增,扩增产物通过Sequence Scanner v1.0导出峰图,Muscle进行序列比对,MEGA 5.1构建进化树,bootstrap 1000检验进化树可靠性,Blat进行同源性分析。结果 纳入本研究MHD患者177例中,抗-HCV阳性13例,感染率为7.3%,HCV-RNA≥1 000U/mL者9例,基因分型以1b型7例(占可分型阳性患者的78%),6a型2例。系统进化树分析显示,3例患者(样本PC1、PC2和PBC17)HCV感染为同一流行株,同源性达99%以上,结合流行病学资料确定1例患者(PC2)为输入性传染源。结论 1b型为深圳市第二人民医院血液透析中心HCV感染的最常见基因型,其次为6a型。通过系统进化树及流行病学分析能可靠地确定HCV感染的同源性,为防治HCV在血液透析中心的医源性传播提供了科学依据。
Objective To examin the genotype distribution and homology analysis of hepatitis C virus (HCV) by the way of phylogenetic tree in the dialysis unit of the second people’s hospital of Shenzhen to provide scientific evidence for preventing HCV infect in hemodialysis unit by homology analysis . Methods The anti-HCV positive dialysis patients were screend for HCV RNA in sera .When the viral load levels were above 1000 U/mL, fragment of HCV 5’UTR and NS5B was severally amplified by nest RT—PCR. The cloned sequences were analyzed using Sequence Scanner v1.0. The genetype of each sample was determined by comparing its nucleotide sequence database .The DNA alignments were generated with Muscle program.The phylogenetic tree was constructed with MEGA 5.1 software using neighbor-joining method and Kimura-two parameter and its reliability was assessed by bootstrap 1000. Homology analysis of DNA alignments was accomplished by Blat software .Result A total of 177 hemodialysis patients were included in this study. Anti-HCV positive are thirteen, accounting for 7.3%. HCV RAN ≥1000 U/mL are 9 samples, in which, genotype 1b are 7 (accounting for 78%), while genotype 6a are 2. Results of polygenetic analysis revealed that three patients (sample PC1、PC2 and PBC17) are the same epidemic strain(homology over 99%)and demonstrated PC2 is the imported infect source with clinical epidemiological data. Conclusion Genotype 1b was the majority in hemodialysis patients with HCV in the second people’s hospital of Shenzhen, while Genotype 6a was secondary in this group. Phylogenetic analysis of NS5B sequences and clinical epidemiological data together provide evidence of infect source and scientific evidence for preventing nosocomial transmission in hemodialysis unit.
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