摘要
目的将特异性抗体免疫富集和聚合酶链反应（PCR）方法有机结合,建立乳制品中阪崎肠杆菌免疫富集及快速检测方法。方法用特异性阪崎肠杆菌单克隆抗体包被的PCR管对样品中病原菌进行特异性免疫富集后,以PCR管中免疫富集的阪崎肠杆菌为模板直接进行PCR扩增,通过特异性、灵敏度、模拟增菌等实验,确认该方法实际检测的可行性并与酶联免疫吸附实验（ELISA）进行比较。结果免疫富集捕获-PCR（IC-PCR）法检测阪崎肠杆菌纯菌液灵敏度可达102~103 CFU/mL,是直接PCR的103~104倍;特异性验证表明,该方法与常见食源性致病菌无交叉反应;模拟带菌实验显示灵敏度可达到103 CFU/mL;模拟增菌实验显示该方法检测阪崎肠杆菌只需增菌6h,而用ELISA试剂盒需要增菌16h。结论该方法灵敏度高、特异性强、检测周期短。
Objective To study a rapid detection method for Enterobacter sakazakii by combining immuno-enrichment with PCR.Methods PCR tubes were coated with monoclonal antibody against Enterobacter sakazakii to capture the suspected bacteria in the samples.PCR was carried out to detect the captured bacteria.The feasibility of the method was assessed with specificity,sensitivity,simulation enrichment experiments and comparison to the results of enzyme linked immunosorbent assay（ELISA）.Results The sensitivity of this method was up to102-103cfu/mL.Compared to the results of direct PCR,the sensitivity of this method was increased by 103-104 times.No positive results was observed for 10 food-borne pathogens other than Enterobacter sakazakii in the tests.The sensitivity of this method in simulation enrichment experiment was up to 103 cfu/mL.The method just needs 6 hours enrichment time,while ELISA method needed16 hours enrichment time.Conclusion The method is sensitivie,specific,and rapid for the detection of Enterobacter sakazakii.
出处
《中国公共卫生》
CAS
CSCD
北大核心
2014年第11期1490-1492,共3页
Chinese Journal of Public Health
