A sandwich-type ELISA for the detection of Listeria monocytogenes using the well-oriented single chain Fv antibody fragment. (September 2017)

• Record Type: Journal Article
• Authors: Liu, Aiping; Xiong, Qing; Shen, Li; Li, Wenli; Zeng, Zhenghai; Li, Cheng; Liu, Shuliang; Liu, Yuntao; Han, Guoquan
• Is Part Of: Food control. Volume 79(2017:Sep.)
• Page Start: 156
• Page End: 161
• Publication Date: 2017-09
• Subjects: Listeria monocytogenes -- Oriented immobilization -- Sandwich-type ELISA -- Single chain Fv antibody fragment; Food -- Quality -- Periodicals; Food -- Analysis -- Periodicals; Food handling -- Periodicals; Food industry and trade -- Quality control -- Periodicals; Aliments -- Industrie et commerce -- Qualité -- Contrôle -- Périodiques; Aliments -- Qualité -- Périodiques; Aliments -- Analyse -- Périodiques; Hygiène alimentaire -- Périodiques; Food -- Analysis; Food handling; Food -- Quality; Periodicals; Electronic journals; 664.07
• DOI: 10.1016/j.foodcont.2017.03.042
• Languages: English
• ISSNs: 0956-7135

Abstract

Abstract: Listeria monocytogenes (LM) poses a serious threat to public health. The present study details the development of a sandwich-type enzyme-linked immunosorbent assay (S-ELISA) for the detection of LM using well-oriented single chain Fv antibody fragment (scFv) and a polyclonal antibody. The anti-LM scFv, used as capture antibody, was gene-optimized and expressed with a C-terminal PS-tag in Escherichia coli BL21(DE3) for oriented-immobilization on hydrophilic polystyrene microtiter plates. The anti-LM rabbit polyclonal antibody, used as the detection antibody, was prepared using heat-killed LM cells as an immunogen. The developed S-ELISA showed great specificity to LM and had a detection limit of lg 6.5 CFU/mL, which was better than the control S-ELISA using non-oriented scFv as the capture antibody. When the S-ELISA was applied to pork and pasteurized milk samples spiked with LM, strong responses were observed. In conclusion, the developed S-ELISA shows great potential for the rapid detection of LM. Highlights: An S-ELISA for rapid detection of LM using well-oriented scFv was developed. Compared to random immobilization, well-oriented scFv can improve sensitivity. The detection limit of LM in enrichment cultures was lg 6.5 CFU/mL. The S-ELISA was applicable for LM detection in pork and pasteurized milk.