维持性血液透析患者丙型肝炎病毒感染基因型及同源性分子流行病学研究  

Molecular epidemiological studies on genotypic and homological distribution of hepatitis C virus in maintenance hemodialysis patients

作　　者：陈嘉[1] 何永成[1] 栾韶东[1] 

机构地区：[1]深圳市第二人民医院肾内科,深圳518000

出　　处：《中国血液净化》2013年第8期425-431,共7页Chinese Journal of Blood Purification

基　　金：广东省科技计划项目立项资助;项目编号:2011B031800035

摘　　要：目的研究深圳市第二人民医院血液透析中心维持性血液透析(maintenance hemodialysis,MHD)患者丙型肝炎病毒(hepatitisCvirus,HCV)感染的基因型及同源性,结合临床流行病学资料,为血液透析中心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阳性的MHD患者的HCV-RNA定量,对HCV-RNA定量≥1×103 IU/ml的MHD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,MEGA5.1构建进化树以确定基因分型,bootstrap1000检验进化树可靠性,Blat进行序列同源性分析。结果 183例MHD患者中,抗-HCV阳性13例,抗-HCV阳性率为7.1%。HCV-RNA定量≥1×103IU/ml者9例中,1b型7例,6a型2例。1b型中,PC1与PC2间的遗传距离为0.000,PBC17与PC2间的遗传距离为0.008,PC29与PBC31间的遗传距离为0.035,PC32与PBC34间的遗传距离为0.102;6a型中,PC11与PC20间的遗传距离为0.068。PC1与PC2的同源性为100%,PC2与PBC17的同源性为99.23%,PC29与PBC31同源性为96.63%,PC32与PBC34的同源性为90.64%,PC11与PC20同源性为93.63%。9例患者的平均感染时间为(67.4±26.3)月,平均透析时间为(81.5±38.1)月,4例(PC2、PC11、PC29和PBC31)为输入性HCV感染者,另5例(PC1、PBC17、PC20、PC32和PC34)是在本中心接受MHD后的8～77月间发现HCV感染。在本中心感染的5例患者中,2例(PC32和PC34)有输血史及更换透析单位,3例(PC1、PBC17、PC20)无输血、更换透析单位、肾移植等感染HCV危险因素,根据流行病学调查,具有同源性的病例PC1和PBC17与感染源(输入性感染者)PC2患者有多次同班或邻班、相邻机位、同一护士操作的血液透析记录。结论深圳市第二人民医院血液透析中心的抗-HCV阳性率为7.1%。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。其中3例1b型患者(样本PC1、PC2和PBC17)HCV感染为同一流行株,同源性达99%以上,结�Objective To determine the genotype distribution and homology of hepatitis C virus （HCV） by using the phylogenetic tree in maintenance hemodialysis （MHD） patients from Shenzhen second people＇ s hospital, in order to provide scientific bases for the prevention of HCV infection in hemodialysis unit. Meth- ods Patients with positive anti-HCV antibodies were examined for HCV RNA in sera. In serum samples with viral ≥1 × 10^3 IU/ml, HCV 5＇ UTR and NS5B fragments were amplified by nested RT-PCR. The sequences of cloned fragments were then analyzed using Sequence Scanner v1.0. Genotype of the fragment was determinedby comparing its nucleotide sequence with the sequences of prototype HCV representing different genotypes in the sequence database. Alignment of sequences was performed using the MUSCLE program. The phyloge- netic tree was constructed with MEGA 5.1 software using neighbor-joining method and Kimura-two parame- ter, and its reliability was assessed by bootstrap 1000. The homology of sequences was analyzed by BLAT software. Results Of the 183 MHD patients enrolled in this study, positive anti-HCV antibodies was found in 13 patients with the HCV infection rate of 7.1%. In the 9 patients with HCV RNA≥1 × 10^3 IU/ml, 7 were in- fected with genotype lb virus and 2 with genotype 6a virus. In the genotype lb samples, genetic distance be- tween PC1 and PC2, between PC2 and PBC17, between PC29 and PBC31, and between PC32 and PC34 was 0.000, 0.008, 0.035 and 0.102, respectively. In the genotype 6a samples, genetic distance between PCI 1 and PC20 was 0.068. Homology between PC1 and PC2, between PC2 and PBC 17, between PC29 and PBC31, be- tween PC32 and PC34, and between PCll and PC20 was 100%, 99.23%, 96.63%, 90.64% and 93.63%, re- spectively. In the 9 patients, the average HCV infection duration was 67.4±26.3 months with the dialysis dura- tion of 81.5±38.1 months. Four patients （sample PC2, PC 11, PC29 and PBC31） were imported infections, and the rest of the 5 patients （sample PC1, PBC17, PC20,

关 键 词：丙型肝炎病毒 基因型 血液透析 系统进化树 

分 类 号：R318.16[医药卫生—生物医学工程]