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酶联免疫吸附测定法用于沙门氏菌一日检测可行性的进一步研究。

Further studies on the feasibility of one-day Salmonella detection by enzyme-linked immunosorbent assay.

作者信息

Wyatt G M, Langley M N, Lee H A, Morgan M R

机构信息

Food Molecular Biochemistry Department, AFRC Institute of Food Research, Norwich Laboratory, Colney, United Kingdom.

出版信息

Appl Environ Microbiol. 1993 May;59(5):1383-90. doi: 10.1128/aem.59.5.1383-1390.1993.

Abstract

A model system previously developed for the rapid detection of Salmonella typhimurium in foods was improved and extended to many other Salmonella serotypes. The original protocol, which consisted of an overnight nonselective culture followed by a specific enzyme-linked immunosorbent assay (ELISA), was modified and improved. A sandwich ELISA which used polyclonal antibodies for the capture stage and a cocktail of monoclonal antibodies for the detector stage was developed. The assay recognized a wide range of Salmonella serotypes; S. enteritidis, the most important serotype in the United Kingdom had a detection limit in the ELISA of about 4 x 10(2) cells ml-1. The cultural stage prior to the ELISA was either a single nonselective broth (incubated for 28 h) or a preenrichment broth (incubated for 7 h) plus a selective broth (incubated for 21 h). Antibodies which bind to cells grown in the unfavorable conditions of a selective medium were selected. It was concluded that, in the future, the shortened protocols for the detection of Salmonella spp. in foods described here will be of considerable value.

摘要

先前开发的用于快速检测食品中鼠伤寒沙门氏菌的模型系统得到了改进,并扩展到许多其他沙门氏菌血清型。原始方案包括过夜非选择性培养,然后进行特定的酶联免疫吸附测定(ELISA),该方案得到了修改和完善。开发了一种夹心ELISA,在捕获阶段使用多克隆抗体,在检测阶段使用单克隆抗体混合物。该测定法可识别多种沙门氏菌血清型;肠炎沙门氏菌是英国最重要的血清型,在ELISA中的检测限约为4×10² 个细胞/毫升。ELISA之前的培养阶段要么是单一非选择性肉汤(培养28小时),要么是预富集肉汤(培养7小时)加选择性肉汤(培养21小时)。选择了与在选择性培养基不利条件下生长的细胞结合的抗体。得出的结论是,未来这里描述的用于检测食品中沙门氏菌属的缩短方案将具有相当大的价值。

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