摘要
为进一步研究外周血单个核细胞 (PBMC)感染丙型肝炎病毒 (HCV)的状况。用逆转录聚合酶链反应 (RT-PCR)、地高辛标记 HCV探针作原位杂交和链酶亲和素 -生物素 (SABC)作免疫组化三项技术 ,检测血清和 PBMC中的 HCV RNA和 HCV NS5 抗原。结果显示 RT- PCR检测的 2 0例慢性丙型肝炎患者 ,除去 4例接受干扰素治疗者 ,余16例中有 14例 (87.5 % )血清 HCV RNA阳性 ,2例血清 HCV RNA阴性者的 PBMC- HCV RNA阳性。对 RT- PCR检测的 9例 PBMC- HCV RNA阳性患者 ,进一步用原位杂交和免疫组化技术 ,分别检测 PBMC中的 HCV RNA和HCV NS5 抗原。显示这几种方法的检测结果是基本一致的 ,呈正相关关系。患者的 PBMC中的 HCV RNA和 HCV NS5抗原呈散在颗粒状或弥漫分布 ,主要位于胞浆中 ,HCV NS5 抗原还分布于胞膜上。证实 HCV可以感染 PBMC,并在其中复制 ,复制部位在胞浆。
To further study the infection of hepatitis C virus(HCV) in peripheral blood mononuclear cells (PBMCs), HCV RNA and HCV NS5 antigen were detected by RT PCR, in situ hybridization with digoxin labled HCV nucleotide probe and strepatadivin biotin enzyme complex assay (SABC) in 20 patients with chronic hepatitis C. The results showed that among the 20 patients with chronic hepatitis C undergoing RT PCR, 14 cases (87.5 %) out of 16 cases were positive for serum HCV RNA and the remaining 2 cases with negative serum HCV RNA were positive for PBMC HCV RNA except 4 patients treated with α interferon were negative for HCV. HCV RNA and HCV NS5 antigen in PBMC were further tested by in situ hybridization and SABC in 9 patients who were HCV RNA positive in PBMC by RT PCR. The results obtained from the three different methods was similar and showed a positive correlation. HCV RNA and HCV NS5 signals in PBMC were granularly or diffusely distributed in cytoplasm. Furthermore, HCV NS5 signal also appeared on the membrane of PBMC. The present data showed that HCV marks determined by RT PCR were credible and confirmed that HCV not only could infect PBMC, but also could be replicated in PBMC cytoplasm.
出处
《同济医科大学学报》
CSCD
2000年第2期174-176,共3页
Acta Universitatis Medicinae Tongji
基金
国家教委启动基金!资助项目 (教留司研 1993- 36 0号 )
