系统进化树在行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 for hepatitis C virus genotype and homology in maintenance hemodialysis patients

作　　者：陈嘉[1] 何永成[1] 栾韶东[1] 丁小强[2] 

机构地区：[1]深圳市第二人民医院肾内科,广东深圳518000 [2]复旦大学附属中山医院肾内科

出　　处：《上海医学》2013年第3期223-228,共6页Shanghai Medical Journal

基　　金：广东省科技计划项目资助(2011B031800035)

摘　　要：目的明确行维持性血液透析(MHD)患者丙型肝炎病毒(HCV)感染的基因型及同源性,为血液透析中心HCV感染的防治提供依据。方法选择1995年1月—2013年2月在深圳市第二人民医院规律行MHD的患者183例,采用荧光定量聚合酶链反应(PCR)测定抗-HCV阳性并行MHD患者的HCV-RNA定量。对HCV-RNA定量≥1×106U/L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 Scannerv1.0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检测进化树的可靠性,Blat进行序列同源性分析,以同源性>95%认为有来自同一传染源的可能。结果 183例行MHD患者中,抗-HCV阳性13例,HCV感染率为7.1%。HCV-RNA定量≥1×106U/L9例:1b型7例(患者分别记录为PC1、PC2、PBC17、PC29、PBC31、PC34、PC32),6a型2例(患者分别记录为PC11、PC20)。1b型中,PC1与PC2间的遗传距离为0.000,PC2与PBC17间的遗传距离为0.008,PC29与PBC31间的遗传距离为0.035;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%,PC11与PC20同源性为93.63%。9例患者的平均感染时间为(7.9±6.1)年,平均透析时间为(7.0±3.1)年。其中,PC1、PC2、PC11、PBC17和PC20均否认有输血史,行透析治疗1～3年间发现HCV感染;PC29、PBC31、PC32和PC34均有输血史;PC2和PC11在入住深圳市第二人民医院血液透析中心前、PC29和PBC31在进行MHD前均已证实为HCV感染。结论 1b型为深圳市第二人民医院血液透析中心行MHD的患者HCV感染最常见的基因型,其次为6a型。PC1、PC2和PBC17的HCV感染为同一流行株,同源性达99%以上,确定PC2为输入性传染源。Objective To examine the genotype distribution and homology of hepatitis C virus （HCV） in patients with maintenance hemodialysis （MHD） and to provide scientific evidence for preventing HCV infection in hemodialysis. Methods The anti-HOV positive dialysis patients were screened for HCV RNA in sera. When the viral load levels were 1 × 106 U/L and more, fragment of HCV 5＇UTR and NS5B were amplified by nest reverse tramscriotion-polymerase chain reaction （RT-PCR）. The cloned sequences were analyzed by using Sequence Scanner vl. 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 by neighbor-joining method and Kimura-two parameter, and its reliability was assessed by Bootstrap 1 000. Homology of DNA alignments was accomplished by Blat software. Results Of the 183 MHD patients enrolled in this, anti-HCV positive results presented in 13 patients, and HCV infection rate was 7. 1 %. There were 9 patients with HCV RNA^I x 106 U/L, including 7 with genotype lb and 2 with genotype 6a. In genotype lb samples, genetic distance between PC1 and PC2, between PC2 and PBC17, between PC29 and PBC31 was 0. 000, 0. 008 and 0. 035, respectively. In genotype 6a samples, genetic distance between PC11 and PC20 was 0.068. Homology between PC1 and PC2, between PC2 and PBC17, between P029 and PBC31, between PC11 and PC20 was 100%, 99.23%, 96.63% and 93.63%, respectively. The nine patients infected HCV for （7.9± 6.1） years dialysis lasted for （7.0± 3. 1） years. Five patients （sample PC1, PC2, POll, PBC17 and PC20） denied transfusion and HCV infection was found during 1 to 3 years of dialysis. Blood transfusion had been performed in 4 patients （P029, PBC31, PC32 and PC3） before dialysis in the Second People＇s Hospital of Shenzhen. HCV infection was proved in 2 patients （PC2 and PC11） before dialysis and in another 2 patients （P029 and PB（331 ）

关 键 词：丙型肝炎病毒 基因型 血液透析 系统进化树 

分 类 号：R459.5[医药卫生—治疗学]