用于检测冷藏牛奶中嗜冷菌的单克隆抗体和间接酶联免疫吸附测定法。

Monoclonal antibodies and an indirect ELISA for detection of psychrotrophic bacteria in refrigerated milk.

作者信息

Gutiérrez R, González I, García T, Carrera E, Sanz B, Hernández P E, Martín R

机构信息

Departamento de Nutrición y Bromatología III, Facultad de Veterinaria, Universidad Complutense, Madrid, Spain.

出版信息

J Food Prot. 1997 Jan;60(1):23-7. doi: 10.4315/0362-028x-60.1.23.

DOI:10.4315/0362-028x-60.1.23

PMID:10465035

Abstract

Monoclonal antibodies generated against live cells of Pseudomonas fluorescens have been used in an indirect ELISA format for the detection of Pseudomonas spp. and related psychrotrophic bacteria in refrigerated milk. The immunorecognition of monoclonal antibodies adsorbed to bacteria bound to the wells of a microtiter plate was performed with rabbit anti-mouse immunoglobulins conjugated to horseradish peroxidase. Subsequent enzymic conversion of the substrate resulted in distinct absorbance differences when assaying milk samples containing psychrotrophic bacteria in the range 10(5) to 10(9) CFU ml(-1) . The detection threshold for the ELISA assay developed in this work is 10(5) CFU ml(-1).

摘要

针对荧光假单胞菌活细胞产生的单克隆抗体已用于间接酶联免疫吸附测定（ELISA）形式，以检测冷藏牛奶中的假单胞菌属及相关嗜冷菌。用与辣根过氧化物酶偶联的兔抗小鼠免疫球蛋白对吸附在微量滴定板孔中结合细菌上的单克隆抗体进行免疫识别。当检测含有10⁵至10⁹CFU/ml范围内嗜冷菌的牛奶样品时，底物的后续酶促转化导致明显的吸光度差异。本研究中开发的ELISA检测方法的检测阈值为10⁵CFU/ml。