Development of a species‐specific monoclonal antibody for rapid detection and identification of foodborne pathogen Vibrio vulnificus

Abstract

Vibrio vulnificus is one of the major cause of foodborne illness related to seafood consumption. This study aimed to develop monoclonal antibodies (MAbs) to enable rapid detection and monitoring food contamination of this pathogen. Five groups of MAbs against V. vulnificus were generated from a mouse immunized with a clinical isolate of V. vulnificus, heat and formalin-inactivated whole cells. The first two groups of MAbs were species-specific to V. vulnificus, recognized all 40 isolates of the bacterium without cross-reactivity to other Vibiro and non-Vibrio species, but bound to different protein antigens. The MAbs in the third group recognized four of nine clinical isolates and two isolates from seafood samples and bound to lipopolysaccharide antigens. The MAbs in the fourth group bound only to all isolates of V. vulnificus and Vibrio alginolyticus, and the MAb in the fifth group was genus-specific bound to all isolates of Vibrio species tested. Based on Dot-enzyme-linked immunosorbent assay, the range of detection sensitivity limits of these MAbs were 10 5 to 10 7 CFU/ml, which is 10-1,000 times less than that of direct PCR detection (10 4 CFU/ml).However, after pre-enrichment of an oyster sample in tryptic soy broth for 6 hr, the detection limit was improved to 1 CFU/ml. This detection sensitivity limit is comparable to that of PCR protocols, which mostly require a pre-enrichment step in sample preparation. The sets of MAbs could provide valuable tools for simple, fast, and low-cost detection of V. vulnificus and Vibrio species in seafood, clinical, and environmental samples. | INTRODUCTIONVibrio vulnificus is a gram-negative, halophilic marine bacterium from the family Vibrionaceae that is widely distributed in estuarine and marine coastal environments. It is frequently isolated from water, sediments, and a variety of seafood, such as shrimp, oysters, fish, and clams (Jones & Oliver, 2009). Since its discovery in 1976 (Hollis, Weaver, Baker, & Thornsberry, 1976), V. vulnificus has become a leading cause of seafood-related deaths throughout the world. Wound infection or gastroenteritis from the ingestion of raw or improperly cooked seafood contaminated with V. vulnificus (especially raw oysters) can cause primary septicemia, particularly in immunocompromised individuals. The death rate is over 50%, and death typically occurs within 72 hr of hospitalization (Oliver, 2005;Yun & Kim, 2018). Therefore, rapid and efficient identification methods are required to facilitate the detection and prevention of V. vulnificus infection from seafood and the environment.The conventional standard microbiological method for identification is based on phenotypic identification, which is time-consuming and labor-intensive. Moreover, several Vibrio species display similar

A Novel Activated Biochar-Based Immunosensor for Rapid Detection of E. coli O157:H7

“…The dot blot test is a widely used method to verify the specific reactivity of antibodies to the desired antigen [ 35 ]. As shown in Figure 6 A, E. coli O157:H7 only reacted to the anti- E. coli pAb by producing black dots.…”

Integration of a CRISPR Cas12a-assisted multicolor biosensor and a micropipette tip enables visible point-of-care testing of foodborne Vibrio vulnificus

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Research progress on detection techniques for point-of-care testing of foodborne pathogens

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