HBOV病毒VP1蛋白的重组表达及在感染检测中的应用

目的]建立检测HBOV感染的免疫学检测方法和进行儿童人群中感染调查。方法]采用PCR扩增克隆HBOV病毒VP1核衣壳蛋白部分基因，将其基因克隆至PET32a，构建原核表达重组质粒，在大肠杆菌中以包涵体形式获得表达。诱导表达后采用凝胶电泳切胶纯化目的蛋白。用重组蛋白作包被抗原建立了间接酶联免疫检测方法。结果]病人血清及献血员血清均以1：50稀释。经检测北京和深圳的85份标本，阳性6人，阳性率7．06％，而正常献血员150人均为阴性，人群中感染率较低。结论]成功重组表达了VP1蛋白，建立了检测抗体的ELISA方法。国内存在该病毒感染，被调查的人群中HBOV感染率较低。

Recombinant expression of VP1 protein of HBOV and its application in detection of infection

Objective] To establish an immunological assay for detection of HBOV infection and investigate the infection of HBOV among children in China. Methods] The fragment of HBOV VP1 gene was amplified by PCR and cloned into PMD - 18T vector for sequencing. After being verified by restricted enzyme digestion, the positive clone was ligated with the prokaryotic expression vector pet32a. The recombinant plasmid was transformed into E, coli BL -21 and the protein was expressed by induction of IPTG, The recombinant protein was purified and used for establishing indirect ELISA for detection of IgG against HBOV. Results] The fusion protein was successfully expressed in inclusion body. The purified protein was coated as antigen to detect serum IgG antibody against HBOV from patients with LRTI by indirect ELISA. The positive rate of IgG antibody against HBOV was 7.06%. The rate of HBOV infection was low in China. Conclusion] The recombinant protein of VP1 of HBOV was expressed successfully and an ELISA for detection of HBOV infection was establishment, This result showed that HBOV infection is low in China.