人巨细胞病毒B基因抗原编码区表达产物的初步应用

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

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(125IproteinA)和非同位素法(碱性磷酸酶标记抗人IgG)检测了已知ELISAHCMV-IgM阳性和阴性血清各77例和4例。结果 同位素法与ELISA一致,非同位素法阳性例为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HCMV52kd糖蛋白抗原主要位于感染细胞核内和细胞表面。结论 HCMV基因工程抗原及其抗体,可检测HCMV早期抗原和抗体,对HCMV感染的临床诊断,鉴别诊断及判断预后有重要意义。

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.