检测产单核细胞李氏杆菌溶血素的胶体金试纸条制备及初步应用

doi:10.11843/j.issn.0366-6964.2022.06.017

Abstract

Abstract: Listeria monocytogenes is an important food-borne pathogen. So it's important and urgent to establish a sensitive and specific method for its rapid detection. The monoclonal antibody of Listeriolysin O (LLO) was screened after prokaryotic expression of recombinant soluble protein His-LLO as antigen, followed by detection of the subtypes, titers and affinity of the antibody. Colloidal gold-based test strip for Listeria monocytogenes LLO was developed via sodium citrate method. Subsequently, the specificity, sensibility, repeatability and stability as well as the primary application of the strip in simulated samples and seafood samples were performed. The results showed that the monoclonal antibody of LLO was specific, stable, repeatable, relatively sensitive and had high affinity, with its affinity constant reaching to 4.25×10⁸ L·mol^-1. The sensibility of the LLO strip in the simulated samples was 3.0×10⁵CFU·mL^-1, which was consistant with that in the pure bacterial cultures. The positive detection of L.monocytogenes from 500 seafood samples was 2.20% (11/500) via LLO colloidal gold-based test strip, which was a bit lower than using standard method (2.40%, 12/500) and PCR detection method (2.40%, 12/500). The coincidence rate of L. monocytogenes positive detection via the aforementioned three methods was 91.67%. Besides, the detection time was deceased to 15 minutes, thereby the efficiency was increased to a greater extent. Therefore, the developed LLO colloidal gold-based test strip could be further used for the rapid detection of L. monocytogenes.

Key words: Listeria monocytogenes, Listeriolysin O, monoclonal antibody, colloidal gold-based test strip, primary application in seafood samples

CLC Number:

S855.1

JIANG Lingli, GAO Youling, HU Xingjuan, ZHANG Xian, YANG Yunkai, WEI Hua, FANG Weihuan. Development and Primary Application of Colloidal Gold-based Test Strip for Rapid Detection of Listeria monocytogenes LLO[J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(6): 1841-1848.

References 20

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Development and Primary Application of Colloidal Gold-based Test Strip for Rapid Detection of Listeria monocytogenes LLO

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