基因芯片对慢性丙型肝炎患者血清病毒基因分型的应用研究

Evaluation of gene chips for sub-genotyping of hepatitis C virus in patients with chronic infection

目的探讨基因芯片对慢性丙型肝炎患者病毒基因分型的应用,分析丙型肝炎病毒(HCV)基因亚型与实验室指标的关系。方法应用基因芯片检测147例HCV-RNA阳性慢性丙型肝炎患者血清病毒基因亚型,并对部分样本进行RT-PCR扩增测序验证;同时检测血清HCV-RNA、丙氨酸氨基转移酶(ALT)、谷氨酸氨基转移酶(AST)、总胆红素(TBil)等相关指标并进行分析。结果 147份HCV患者血清样本中1b型120例(81.6%);2a型13例(8.8%);3a型1例(0.7%),6a型4例(2.7%),未分型9例(6.1%);基因芯片对基因亚型检出率为93.9%,分型结果与测序符合率100%。1b型患者与非1b型患者比较,HCV-RNA、ALT、AST、TBil等各指标差异均无统计学意义(P均>0.05)。结论基因芯片技术适合对临床样本丙肝病毒基因亚型的快速检测,本地区慢性丙肝患者病毒基因亚型以1b型为主,其防治工作值得引起相当重视。

Objective To evaluate the potential use of gene chips for sub-genotyping of hepatitis C virus（HCV） in patients with chronic infection and to analyze the correlation between HCV subgenotypes and serum markers.Methods DNA microarray was used to determine HCV subgenotypes in 147 patients with chronic infection,and HCV RNA of some blood samples was amplified by RT-PCR and sequenced for verification.Meanwhile levels of liver function related serum markers were also measured,such as ALT,AST and TBil.Results Out of total 147 blood samples,138 were genotyped successfully by gene chip with detection rate of 93.9%,which included 120 of subgenotype 1b（81.6 %）,13 of 2a（8.8%）,1 of 3a（0.7%） and 4 of 6a（2.7%）.They were all further verified by DNA sequencing with 100% accuracy.There was no significant difference of liver function indicators as ALT,AST and TBil between 1b and non-1b subgenotype patients（P〉0.05）.Conclusions The gene chip technology is a fast,convenient and accurate method for HCV subgenotyping of clinical samples.The subgenotype 1b is found to be the dominant one over the area.