Development of rapid and sensitive antiflagellar monoclonal antibody based lateral flow device for the detection of Vibrio vulnificus from oyster homogenate

Abstract

The current study was designed to develop an antibody based chromatographic lateral flow detection device (LFD) for rapid identification of Vibrio vulnificus from oyster homogenate. Anti-H monoclonal antibodies against V. vulnificus were prepared and used in the development of the LFD device. The LFD device was tested for its specificity, sensitivity and storage stability. The LFDs were tested against seven different Vibrio spp. along with three different V. vulnificus isolates. All Vibrio except for V. vulnificus failed to produce a visible signal on the device. The LFD was able to detect the presence of V. vulnificus at levels of approximately 4 log CFU/ml from phosphate buffered saline and oyster homogenate without any pre-enrichment process. When combined with 6.66 h enrichment period the LFD was able to detect pathogen counts as low as 10 CFU/ml from oyster homogenate. The storage stability and effects of temperature abuse on the LFD were determined by storing LFD at 4 °C, 25 °C and 40 °C for two weeks. It was found that two weeks storage in extreme temperatures, 4 °C and 40 °C, did not affect the sensitivity of the LFD. The results indicate that LFD were able to identify target pathogens from oyster homogenate accurately and could serve as rapid and sensitive V. vulnificus screening method from oysters with or without enrichment.