人偏肺病毒表面糖蛋白G的原核表达及哈尔滨某医院非呼吸道感染住院患者血清学检测  

Human metapneumovirus surface glycoprotein G prokaryotic expression and non-respiratory tract infection in hospitalized patients with serological testing in a hospital in Harbin

作　　者：喻欢[1] 赵景波[1] 武旭芳[2] 马玲[1] 王永[1] 

机构地区：[1]哈尔滨医科大学公共卫生学院流行病学教研室,黑龙江哈尔滨150081 [2]内蒙古医科大学药学院,内蒙古呼和浩特010010

出　　处：《中华疾病控制杂志》2013年第6期476-479,共4页Chinese Journal of Disease Control & Prevention

基　　金：国家自然科学基金(30800984)

摘　　要：目的构建人偏肺病毒(human metapneumovirus,hMPV)两种基因型的表面糖蛋白G基因原核表达体系,以成功表达的hMPV表面糖蛋白为基础,建立稳定有效的血清抗体检测方法。方法分别以hMPV两种基因型的G基因质粒为模版扩增目的片段,克隆到pBluescriptⅡPhagemid载体,测序后,将目的片段经双酶切后克隆到表达载体pET 32-a-(c+),诱导表达目的蛋白,作为包被抗原,用ELISA方法对106份血清样品进行检测。结果成功表达了hMPV两种基因型的表面糖蛋白G,并用此检测哈尔滨地区因非呼吸道感染住院患者血清样品106份,在不同年龄组中,抗hMPV-A型的血清阳性率分别为:≤10岁6.25%、10～20岁6.67%、20～40岁12.50%、40～60岁16.67%、>60岁21.74%;抗hMPV-B型的血清阳性率分别为:≤10岁12.50%、10～20岁6.67%、20～40岁18.75%、40～60岁22.22%、>60岁30.43%。结论成功表达了两种基因型hMPV的表面糖蛋白G,并建立了检测人血清抗体的ELISA方法。检测结果显示hMPV感染在哈尔滨地区人群中不少见,整体上来说,随着年龄的增加,血清抗体阳性率也是增高的。Objective To construct the two genotypes of human metapneumovirus surface glycoprotein G gene prokaryotic expression system,and to establish a stable and effective serum antibody detection method based on the successful expression of hMPV surface glycoprotein.Methods G gene was amplified from hMPV two genotypes plasmid,then cloned into the pBluescript Ⅱ Phagemid vector and confirmed by sequencing.After dual enzyme,target fragments were subcloned into the pET 32-a-（c ＋） vector and then expressed.The expressed target protein could be used as a coating antigen to detect 106 serum samples by ELISA.Results The two genotypes of hMPV surface glycoprotein G were expressed successfully and they were used to detect serum samples of non-respiratory infection hospitalized patients,the seroprevalence of antihMPV-A type was： 6.25%（≤10 years）;6.67%（10-20 years）;12.50%（20-40 years）;16.67%（40-60 years）;21.74%（ 60 years） respectively.The seroprevalence of anti-hMPV-B-type was： 12.50%（≤10 years）;6.67%（1020 years）;18.75%（20-40 years）;22.22%（40-60 years）;30.43%（ 60 years） respectively.Conclusions This study successfully expresses two genotypes of hMPV surface glycoprotein G and establishes the ELISA method for the detection of human serum antibodies.The ELISA results show that the hMPV infection is common in Harbin,and the seroprevalence totally increases following age.

关 键 词：病毒 血清学实验 酶联免疫吸附测定 

分 类 号：R181.3[医药卫生—流行病学]