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本文采用丙型肝炎病毒(HCV)基因中高度保守的5′端非编码区的两对引物,建立了检测 HCV 核酸(HCV-RNA)的巢式(两步法)多聚酶链反应(PCR),对其中标本 RNA的提取、互补 DNA(cDNA)合成时引物的浓度、PCR 的反应体积以及 PCR 时温度循环程序等进行了优化选择。用该法检测了67例非甲非乙型肝炎患者的血清标本,发现52例(78%)为HCV-RNA 阳性。在丙型肝炎病毒抗体(抗-HCV)阴性的27例非甲非乙型肝炎患者中检出16例(59%)HCV-RNA 阳性。这些结果再次证实 HCV 感染是非甲非乙型肝炎的重要病因,并提示仅检测抗-HCV 可能会低估 HCV 的感染率。
In this paper, two pairs of primers of highly conserved 5’-end non-coding region of hepatitis C virus (HCV) gene were used to establish a nested (two-step) polymerase chain reaction (PCR) The extraction of the sample RNA, the concentration of the complementary DNA (cDNA) synthesis primer, the reaction volume of the PCR, and the PCR temperature cycling program were optimized and selected. Serum samples from 67 patients with non-A, non-B hepatitis were detected by this method and 52 (78%) were positive for HCV-RNA. Sixteen (59%) HCV-RNA positive were detected in 27 non-A, non-B hepatitis patients with negative HCV antibody (anti-HCV). These results again confirm that HCV infection is a significant cause of non-A, non-B hepatitis and suggest that testing for anti-HCV may underestimate the rate of HCV infection.