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免疫磁珠法与表面黏附免疫荧光技术用于快速检测肉类中单核细胞增生李斯特菌和无害李斯特菌的比较

A comparison of immunomagnetic and surface adhesion immunofluorescent techniques for the rapid detection of Listeria monocytogenes and Listeria innocua in meat.

作者信息

Duffy G, Sheridan J J, Hofstra H, McDowell D A, Blair I S

机构信息

National Food Centre, Dublin, Ireland, UK.

出版信息

Lett Appl Microbiol. 1997 Jun;24(6):445-50. doi: 10.1046/j.1472-765x.1997.00139.x.

Abstract

An immunomagnetic immunofluorescent method was investigated for the rapid detection of Listeria monocytogenes and Listeria innouca. This technique involved enrichment of the suspect sample at 30 degrees C overnight. Listeria monocytogenes cells were isolated from the enriched sample using immunomagnetic separation and Listeria were subsequently visualized using an immunofluorescent microscopy technique. This technique was used in the detection of Listeria cells from pure culture, inoculated beef mince samples and naturally contaminated retail beef mince samples. A detection level of approximately 1 x 10(3) cfu ml-1 was achieved. When compared with traditional detection methods no false negatives or positives were recorded for L. monocytogenes or L. innocua. The immunomagnetic immunofluorescent technique had a detection level similar to a previously described surface adhesion immunofluorescent technique. Isolation of the Listeria cells by surface adhesion involved dipping a membrane attached to a microscope slide into the enriched sample for 10 min. This was quicker and simpler to perform than the immunomagnetic separation technique which took 2 h to carry out.

摘要

研究了一种免疫磁珠免疫荧光法用于快速检测单核细胞增生李斯特菌和伊氏李斯特菌。该技术包括在30℃下对可疑样品进行过夜富集。使用免疫磁珠分离从富集样品中分离出单核细胞增生李斯特菌细胞,随后使用免疫荧光显微镜技术对李斯特菌进行可视化观察。该技术用于检测纯培养物、接种碎牛肉样品和天然污染的零售碎牛肉样品中的李斯特菌细胞。实现了约1×10³ cfu ml⁻¹的检测水平。与传统检测方法相比,单核细胞增生李斯特菌或无害李斯特菌未记录到假阴性或假阳性结果。免疫磁珠免疫荧光技术的检测水平与先前描述的表面粘附免疫荧光技术相似。通过表面粘附分离李斯特菌细胞的方法是将附着在显微镜载玻片上的膜浸入富集样品中10分钟。这比需要2小时才能完成的免疫磁珠分离技术更快、更简单。

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