DNA微列阵技术检测戊型肝炎病毒方法的建立及初步应用

目的建立DNA微列阵技术检测戊型肝炎病毒方法,进而探讨该方法用于检测临床标本的可行性。方法通过生物医学数据库戊型肝炎病毒基因进行检索和筛选,应用分子生物学软件,进行序列分析、引物及探针设计,并进行验证,建立了戊型肝炎病毒DNA微列阵技术检测方法。结果试验所设计的探针仅与HEV的PCR产物杂交呈阳性,与乙肝、丙肝等对照病毒的PCR产物杂交呈阴性。敏感性试验显示,该方法比同巢式PCR方法检测戊型肝炎病毒敏感度要高,用该方法检测了长春地区50份疑似HEV临床病料,38份阳性;而用巢式PCR法扩增HEV ORF1基因确诊为阳性的只有35份。结论利用DNA微列阵技术检测戊型肝炎病毒方法,特异性和敏感性强,可作为HEV临床标本检测方法。

DNA microarray detection of hepatitis E virus Establishment and preliminary application of methods

Objective To establish a hepatitis E virus nucleic acid probe detection methods,and then explore the method used to detect the feasibility of clinical samples.Methods Biomedical hepatitis E virus genome database retrieval and filtering,the application of molecular biology software for sequence analysis,primer and probe design and carry out verification,establishment of hepatitis E virus nucleic acid probe detection methods.Results The results of tests designed to probe the PCR product only with HEV positive hybridization with hepatitis B,hepatitis C virus,such as the control PCR product of hybridization were negative.Sensitivity tests have shown that this method is compared with the nested-PCR detection of hepatitis E virus,And Using this method to detect 50 copies in Changchun area of clinical disease suspected HEV material,38 were positive;while nested-PCR amplification of HEV ORF1 gene diagnosed as positive,only 35 copies.Conclusion The use of hepatitis E virus nucleic acid probe detection methods,specificity and sensitivity of strong clinical samples can be used as HEV detection methods.