EB病毒DNA聚合酶基因的表达与纯化鼻咽癌病人诊断中的应用

以Ｅｐｓｔｅｉｎ－Ｂａｒｒ病毒（ＥＢＶ）ＤＮＡ聚合酶为抗原,建立了简便、快速、敏感和特异的鼻咽癌诊断方法。构建原表达载体ｐＲＳＥＴ－ＤＮＡ聚合酶及其亚克隆ＰＲＳＥＴ－Ａ１和ＢＬ２１（ＤＥ３）中表达的产物,经Ｗｅｓｔｅｒｎ－ｂｌｏｔ检测其抗原性并用于检测鼻咽癌（ｎａｓｏｐｈａｒｙｎｇｅａｌ ｃａｒｃｉｎｏｍａ,ＮＰＣ）病人血清中的抗体。在ＤＰＣ病人血清中存在抗ＥＢ病毒ＤＮＡ聚合酶的ＩｇＧ抗体,并证明

Study on the Expression and Purification of Epstein-Barr Virus DNA Polymerase and Its Application to Diagnosis of Nasopharyngeal Carcinoma

In order to establish a simple, rapid, sensitive and specific assay for diagnosis of nasopharyngeal carcinoma (NPC), we prepard Epstein-Barr virus (EBV) DNA polymerase antigen by genetic engineering. DNA polymerase was expressed in BL21(DE3) using pRSET vector. The expressed product was characterized by SDS-PAGE and Western-blot. The IgG antibody angainst DNA polymerase of EBV in sera of NPC patients can be testified by expressed DNA polymerase through Western-blot and ELISA. The DNA polymerase is divided into the front 1/3 and the last 2/3 A2 parts, on A2 the antigenicity of DNA ploymerase locates. The sensitivity and specificity of Western-blot for detecting A2/IgG antibody was 80% and 100% respectively. The ELISA test established to detect A2/IgG antibody in sera of 46 NPC patients and 46 non-NPC patients also showed high sensitivity (89%) and specificity (93%).