人巨细胞病毒B基因抗原编码区表达产物的初步应用

目的 人巨细胞病毒（ＨＣＭＶ）５２kd糖蛋白重组表达质粒（pＨＣＭＶ５２）经诱导、 增殖、提取并纯化表达产物。方法 采用同位素法（＾１２５Ｉprotein Ａ）和非同位素 法（碱性磷酸酶标记抗人ＩgＧ）检测了已知ＥＬＥＳＡＨＣＭＶ－ＩgＭ阳性和阴性血清各７７例和４例。结果 同位素汪与ＥＬＩＳＡ一致，非同位素汪阳性例为７５/７７例。表 达产物免疫家兔制备的抗血清，用ＡＢＣ法检测肝组织中ＨＣＭＶ抗原，发现ＨＣＭＶ５２

Preiliminary use of expression product of HCMV B gene coding region in pathlogy and clinics

Objective To make preliminary use of the expression product of HCMV B gene coding region in pathology and clinics.Methods A recombinant expression plasmid(pHCMV 52)containig human cytomegalovirus B gene sequence coding for 52kda antigen was constructed,transformed into E coli,and then induced,amplified and extracted.The purified recombinant antigen and antibodies to the recombinant antigen were prepared.The Dot blot and Western transfer showed that the protein expressed from the cloned gene and its antibodies had specifically immunologic activities.77 cases of HCMV IgM ELISA postive sera and 4 cases of negative sera were detected by using radiolabeling( 125 I Protein A)and nonradiolebeling(calf alkaline phosphatase labeled antibodies to human IgG).Also used were the antibodies to the recombinant antigen detected with avidin biotin peroxidase complex(ABC)technique.Results The positive ratio of 125 I Protein A method was 100% and that of nonradiola beling 97%(75 cases).HCMV 52kda glycoprotein was mainly located in the nuclei and on the membrane of the HCMV infected hepatocytes.Conclusion The authors suggest that the application of the expression product of HCMVgene be of clinical significance in the diagnosis and prognostication of HCMV infection.