丙型肝炎病毒干扰素敏感决定区聚合酶链反应检测方法的建立及应用  

Establishment and application of polymerase chain reaction method for detection of interferon sensitivity determining region of hepatitis Cvirus

作　　者：张剑平[1] 徐道振[1] 闫杰[1] 何忠平[2] 董庆鸣[1] 戴旺苏[1] 沈冰[1] 魏红山[1] 成军[1] 

机构地区：[1]北京地坛医院传染病研究所,北京市100011 [2]北京佑安医院检验科,北京市100054

出　　处：《世界华人消化杂志》2005年第24期2874-2876,共3页World Chinese Journal of Digestology

摘　　要：目的:建立一种以丙型肝炎病毒(HCV)干扰素敏感决定区(ISDR)特异性引物为基础的聚合酶链反应(PCR)方法,对HCVISDR进行序列测定.方法:根据GenBank中6株HCV全序列核苷酸序列,设计共同的外引物及三组ISDR型特异性内引物.建立了HCVISDRPCR检测方法,对27例HCV基因型1b的慢性丙型肝炎患者血清HCVRNA进行测定,并对PCR阳性标本的产物进行多位点分析.结果:A组PCR结果阳性率为37%(10/27),A-B组为30%(8/27),A-C组为4%(1/27),A-B-C组为22%(6/27),三组均阳性者有5例为使用IFN治疗6-12mo后定性PCR检测HCVRNA转阴,停药3mo后PCR检测HCVRNA结果仍为阴性.结论:以ISDR特异性引物为基础的PCR法可靠、简便,可用于丙型病毒性肝炎患者对HCV进行敏感决定区的测定,用于以NS5A-ISDR突变数量来预测HCV基因型1b感染的患者对IFN的持续应答.AIM： To establish a polymerase chain reaction （PCR） method based on the special primers of the interferon sensitivity determining region （ISDR） of hepatitis C virus （HCV）, and to detect the sequence of the HCV ISDR. METHODS： The universal outer primers and 3 suits of ISDR type-specific inner primers （A, B, C group） were designed on the basis of the nucleotide sequences of 6 typical full-genomic HCV. The HCV RNA in the serum that colleted from 27 patients infected with chronic HCV genotype lb was tested by the established PCR method, and multi-site analysis was performed on the PCR positive products. RESULTS： The positive rats were 37% （10/27）, 30% （8/27）, 4% （1/27）, and 22% （6/27） in group A, group A-B, group A-C, and group A-B-C after detection by the established PCR method. For the 6 HCV RNA positive patients, HCV RNA became negative when they were treated with IFN for 6 to 12 mo, and HCV RNAwas still negative in those patients 3 mo after the treatment. CONCLUSION： The PCR method based on the ISDR of HCV is simple and reliable, and can be used in the detection of HCV ISDR in patients with hepatitis C and in the prediction of the persistent response to the IFN in HCV genotype lb infected patients by the number of mutations within the ISDR.

关 键 词：丙型肝炎病毒 干扰素敏感区 聚合酶链反应 

分 类 号：R450[医药卫生—治疗学]