应用磁免疫技术快速检测三种新发病原菌

Rapid detection of three emerging pathogens by new methods based on Immunomagnetic separation

目的应用磁免疫分离技术（IMS）建立对3种新发病原菌（O139群霍乱弧菌、EHEC O157∶H7、单增李斯特菌）的快速检测方法。方法将IMS分别与显色培养（chromogenic media,Ch）和荧光定量PCR技术结合,建立了IMS-Ch、IMS-PCR两种新方法,研究不同方法的灵敏性和特异性,并对高危样品的检测效果进行评价。结果 IMS-Ch法24 h能检出目标菌含量≥50 cfu/10 g的样品,IMS-PCR法用于检测EHEC O157∶H7和霍乱弧菌,9 h可达检出限为10~20 cfu/10 g,实际检测效果评价的尤等指数〉95%。结论 IMS-PCR是一种可信的快速筛选方法,IMS-Ch法为较国标法灵敏、快速的确证方法。

Objective To develop two rapid assays to detect three emerging pathogens（O139 Vibrio cholerae,EHECO157∶H7,Listeria monocytogenes）by using immunomagnetic separation（IMS）.Methods Two assays of IMS-Ch and IMS-PCR were established in combination with chromogenic media（Ch） and quantitative fluorescencet PCR.The sensitivity and specificity of two assays were compared and the evaluation on the effectiveness on the detections of high risk samples was conducted.Results IMS-Ch could detect the samples containing ≥ 50 cfu/10 g pathogen in 24 hours and the detection threshold of IMS-PCR to EHEC O157∶ H7 and Vibrio cholerae was 10-20 cfu/10 g in 9 hours.The Youden＇s index of this assay was more than 95%.Conclusion IMS-PCR is a reliable and rapid screening assay,IMS-Ch is a more sensitive and rapid confirmation assay.