肝细胞中人巨细胞病毒抗原的免疫组化检测

Immunohistochemical Detection of Antigen of Human Cytomegalovirus in Hepatocyte

以人巨细胞病毒（ＨＣＭＶ）５２ｋｄ糖蛋白表达质粒（ｐＨＣＭＶ５２）扩增后提取的表达产物免疫家兔制备抗血清，采用生物素标记的免疫组化法（ＡＢＣ法）检测９例人体肝组织中的ＨＣＭＶ５２ｋｄ糖蛋白抗原。结果：临床诊断为巨细胞包涵体病的８例中７例为阳性，其ＨＣＭＶ５２ｋｄ糖蛋白主要位于肝细胞核内和细胞膜上。ＡＢＣ法方法简便、敏感性强及特异性高，且能直接判断组织中是否存在ＨＣＭＶ抗原并定位，对协助临床诊断、鉴别诊断和判断预后均具有重要意义。

The expression plasmid of human cy-tomegalovirus(HCMV)52kd glycoprotein was in-duced and amplified, and the recombinant protein wasextracted.The rabbit antiserum to the recombinantprotein was prepared.The antiserum reacted to the antigen of HCMV AD169, and the titer of antiserumwas 1: 3200 by using125 I-Protein A. Using Avidin-Biotinperoxidase complex technique(ABC method),we detected the HCMV 52 kd glycoprotein in thehapatic slides of 9 cases,Among them,one case(aninfant died of congenital heart disease)was held as anegative control.The other 8 cases(patients withclinically diagnosed cytomegalic inclusion disease) had cation.viral hepatitis histopathologically.of these 8 cases,7had positive signals but found in the nucleus and cy-tomembrane of the hepatocytes,no positive signal washund in the hepatocytes of the control. The resultshowed that HCMV 52kd glycoprotein was located inthe nucleus and on the membrane of the infected hepa-tocyte. The antibody prepared with the antigen ofgene engineering could find the virus antigen and rep-resent the virus gene state which was active or silent.ABC methdod is not only simple,sensitive and specific,but also effective in localization. It is of importance toclinical diagnosis, differential diagnosls and prognosti-cation.