应用基因重组抗原诊断丙型肝炎研究

丙型肝炎病毒(Hepatis C virus,HCV)感染者的血液中病毒含量极低,还没有适当的方法可以直接检测病毒。现在常用免疫学方法测定特异性抗体[1]或利用 PCR 技术检测病毒核酸。由于 HCV基因组变异高达33%,PCR检测易发生假阳性和假阴性。同时由于丙肝抗原高度的变异性[2],虽然目前ELISA方法应用广泛,但是单一的诊断抗原已不能满足临床要求,需要多价抗原联合应用作为诊断试剂。目前,在我国流行的丙型肝炎病毒主要是Ⅱ型和Ⅲ型,国外的丙型肝炎诊断试剂研制单位主要是以Ⅰ型为主[3],而且进口的丙型肝炎诊断试剂昂贵,不适合在我国大量使…

The Diagnostic Research of Hepatitis C with Recombinant Antigen

In order to improve the diagnostic efficiency of the hepatitis C in China, the DNA fragments of core, C33c,NS4 in Hepatitis C virus(HCV) were connected and subcloned to pET24a( ). The pET24a-CCN was highly expressed with the inducement IPTG in the E.coli BL21(DE3). In 1L bacterial culture medium,30mg purified recombinant proteins were available using His Binding Resin under denatured condition. Used as diagnostic antigen to detect the human hepatitis C by ELISA, The results showed that the 100% positive coincidence rate (66/66) and 94.5% negative coincidence rate (86/91) were obtained. To detect 46 positive HAV sera and 46 positive HBs sera, all the results were negative. At the same time, the freeze-dried recombinant antigen detects 66 HCV positive sera, all the results were positive. Therefore, the recombinant antigen was sensitivity, specialty and stability to detect hepatitis C. It was an ideal reagent to detect hepatitis C.