基于针对食品中单核细胞增生李斯特菌的单克隆抗体的抗原捕获酶联免疫吸附测定法的开发与验证

Development and validation of an antigen capture ELISA based on monoclonal antibodies specific for Listeria monocytogenes in food.

作者信息

Portanti Ottavio, Di Febo Tiziana, Luciani Mirella, Pompilii Cinzia, Lelli Rossella, Semprini Primula

机构信息

Istituto Zooprofilattico Sperimentale dell'Abruzzo e del Molise G. Caporale, Campo Boario, 64100 Teramo, Italy.

出版信息

Vet Ital. 2011 Jul-Sep;47(3):281-90, 271-80.

PMID:21947967

Abstract

A capture enzyme-linked immunosorbent assay (ELISA) for the identification of Listeria monocytogenes in food was standardised and validated. The assay was refined by analysing samples of meat, seafood, dairy products, pasta and flour. The method was found to be 100% specific for Listeria spp. tested, with a limit of sensitivity of 6.6 × 10(3) colony-forming units (cfu)/ml. Comparison of L. monocytogenes capture ELISA against the official International Organization for Standardization (ISO) method 11290-1:1996 for the isolation and identification of L. monocytogenes in food matrices produced a significant concordance index. The assay was validated on food matrices including meat, seafood and dairy products in line with ISO 16140:2003 concerning qualitative analytical methods. The assay was found to be accurate, specific, sensitive, selective, reproducible and fast, resulting in lower costs and faster turnaround in microbiological screening of foods.

摘要

一种用于鉴定食品中单核细胞增生李斯特菌的捕获酶联免疫吸附测定（ELISA）方法得到了标准化和验证。通过分析肉类、海鲜、乳制品、面食和面粉样本对该测定方法进行了优化。结果发现该方法对所检测的李斯特菌属具有100%的特异性，灵敏度限值为6.6×10(3) 菌落形成单位（cfu）/毫升。将单核细胞增生李斯特菌捕获ELISA与官方国际标准化组织（ISO）11290-1:1996方法（用于食品基质中单核细胞增生李斯特菌的分离和鉴定）进行比较，得出了显著的一致性指数。根据ISO 16140:2003关于定性分析方法的标准，该测定方法在包括肉类、海鲜和乳制品在内的食品基质上进行了验证。结果发现该测定方法准确、特异、灵敏、有选择性、可重复且快速，从而降低了食品微生物筛查的成本并缩短了周转时间。