【摘要】 目的建立快速检测土壤中类鼻疽伯霍尔德杆菌(B.pseudomallei)的双抗体夹心酶联免疫吸附测定(ELISA)法,用于该菌疫源地的流行病学调查。方法通过基因工程克隆并表达B.pseudomallei的特异性鞭毛蛋白,纯化后免疫小鼠,制备针对特异性B.pseudomallei鞭毛蛋白的单克隆抗体。以单克隆抗体为捕获抗体,全菌体多克隆抗体为检测抗体,建立双抗体夹心ELISA方法。结果成功建立双抗夹心ELISA法,采用该法对土壤中细菌进行检测,其敏感性和特异性分别为100%(15/15)和97.5%(40/41);对广西地区154个土壤样品进行检测,12个土壤样品为阳性。结论双抗体夹心ELISA法对土壤中B.pseudomallei检测的敏感性和特异性好,可用于类鼻疽的流行病学调查。更多还原

【Abstract】 Objective To establish double-antibody sandwich enzyme-linked immunosorbent assay(ELISA)for rapid detection of soil melioidosis Burkholderia pseudomallei(B.pseudomallei)and epidemiological investigation of this bacteria in epidemic focus.Methods Specific B.pseudomallei flagellin was cloned and expressed through genetic engineering.It was used to immunize mice after purification in order to prepare monoclonal antibody against specific B.pseudomallei flagellin.Double-antibody sandwich ELISA method was established by monoclonal antibody being served as capture antibody and polyclonal antibody against whole B.pseudomallei as detection antibody.Results Double-antibody sandwich ELISA method was successfully established.It was then employed to detect bacteria in soil.It’s detective sensitivity and specificity were 100%(15/15)and 97.5%(40/41),respectively,and 12 soil samples were found positive when 154 soil samples obtained from Guangxi region were undergone bacteria detection.Conclusion Double-antibody sandwich ELISA method possesses high sensitivity and specificity in detection of soil B.pseudomallei,and can be used in epidemiological investigation of melioidosis.更多还原