A multiplex PCR for the detection of Vibrio vulnificus hazardous to human and/or animal health from seafood

Roig, Arnau Pérez; Salido, Héctor Carmona; Sanjuán, Eva; Fouz, Belén; Amaro, Carmen

doi:10.1016/j.ijfoodmicro.2022.109778

Cited by 12 publications

(10 citation statements)

References 26 publications

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“…Laboratory samples indicated that all samples were appropriately confirmed at 100%. Comparing our results to those in the literature showed that they were about the same or better in terms of specificity, sensitivity, and cost (Denyingyhot et al, 2022;Edwards & Gibbs, 1994;Galal-Khallaf, 2021;Lourenço et al, 2022;Roig et al, 2022;Sangthong et al, 2021;Yanaso et al, 2021;Zhao et al, 2021).…”

Section: Resultssupporting

confidence: 72%

Multiplex Analysis of Animal Species Origins in Feedstuffs and Foodstuffs for the Prevention, Control and Eradication of Transmissible Spongiform Encephalopathies (TSEs) by PCR

Safdar¹,

Özaslan²

2022

EPHELS

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Recently, highly strict regulations for the prevention, control, and elimination of transmissible spongiform encephalopathies (TSEs) were put in place by the European Parliament and the Council of the European Union. Furthermore, because of health concerns, there is a greater demand for knowledge about the composition of feeds and foods, particularly pet food and ruminant feed, so determining the species of origin is critical. Therefore, there is a dire need for an advanced technique that should be rapid, specific, and inexpensive. The purpose of this study was to develop an M-PCR (multiplex PCR) assay that could identify TSEs causing origin species concurrently in food and feed products while employing fluorescent dyes that were less expensive than doubly labelled probes. The development and optimization of M-PCR allowed for the analysis of the origin of animal species' DNAs in complicated feed and food matrices. The primers were designed using specific segments of DNA sequences of the targeted species (bovine: 271bp, ovine: 119bp, caprine: 224bp). The optimized M-PCR assay may be a useful tool for confirming the species origin of feedstuffs and commodities subjected to denaturing technologies, according to the assay's results.

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Section: Resultssupporting

confidence: 72%

Multiplex Analysis of Animal Species Origins in Feedstuffs and Foodstuffs for the Prevention, Control and Eradication of Transmissible Spongiform Encephalopathies (TSEs) by PCR

Safdar¹,

Özaslan²

2022

EPHELS

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“…In addition, the sensitive detection of V. vulnificus in oysters was realized by RAA-TS-DTL. As shown in Table 5, the classical qPCR, multiple PCR and droplet digital PCR (ddPCR) methods for V. vulnificus had LODs ranging from 10 to 100 CFU/mL [8,22,23]. The RAA-TS-DTL developed on the basis of the gyrB gene could detect V. vulnificus in oyster at levels as low as 6 CFU/mL, which was 15 times more sensitive than that of the qPCR method established by D'Souza et al (100 CFU/mL) [8].…”

Section: Discussionmentioning

confidence: 99%

A Novel RAA Combined Test Strip Method Based on Dual Gene Targets for Pathogenic Vibrio vulnificus in Aquatic Products

Liu,

Zhang,

et al. 2023

Foods

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Vibrio vulnificus can cause disease in aquatic animals and humans, therefore, rapid and simple field detection of pathogenic V. vulnificus is important for early disease prevention. In this study, a novel recombinase-aided amplification (RAA) combined test strip with double T-lines (RAA-TS-DTL) was developed for the rapid detection of V. vulnificus in aquatic products. Pathogenic V. vulnificus was detected using the virulence vvhA gene and the housekeeping gene gyrB gene as the dual target of the test strip. The RAA-TS-DTL method showed 100% specificity for V. vulnificus, and no cross-reaction was observed with Vibrio spp. or other bacteria (n = 14). Furthermore, sensitive detection of V. vulnificus in oysters was achieved. The LODs of the gyrB and vvhA genes were 6 CFU/mL and 23 CFU/mL, respectively, which was about five times higher than that of the commercial test strip. The method was validated with spiked samples (n = 60) of fish, shrimp and oyster. The consistency between RAA-TS-DTL and the traditional culture method was 97.9%. In addition, the entire process of detection, including preparation of the sample, could be completed within 50 min. Our results indicated that the developed RAA-TS-DTL was a reliable and useful tool for rapid screening or on-site detection of pathogenic V. vulnificus in aquatic products and aquaculture water.

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“…Usually the thermostable direct hemolysin ( tdh ) and thermostable related hemolysin ( trh ) are used as PCR targets [ 102 , 103 ]. Lastly, V. vulnificus produces hemolysins (hemolysin A can by researched through targeting the vvha gene), proteases and cytotoxins, contributing to its pathogenic nature and ability to cause tissue destruction [ 104 ]. Additionally, colistin resistance has been identified in STEC and Salmonella isolated from shellfish [ 105 ].…”

Section: How To Translate Microbial Analysis In Term Of Public Health...mentioning

confidence: 99%

A Comprehensive Review for the Surveillance of Human Pathogenic Microorganisms in Shellfish

Desdouits,

Reynaud,

Philippe

et al. 2023

Microorganisms

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Bivalve molluscan shellfish have been consumed for centuries. Being filter feeders, they may bioaccumulate some microorganisms present in coastal water, either naturally or through the discharge of human or animal sewage. Despite regulations set up to avoid microbiological contamination in shellfish, human outbreaks still occur. After providing an overview showing their implication in disease, this review aims to highlight the diversity of the bacteria or enteric viruses detected in shellfish species, including emerging pathogens. After a critical discussion of the available methods and their limitations, we address the interest of technological developments using genomics to anticipate the emergence of pathogens. In the coming years, further research needs to be performed and methods need to be developed in order to design the future of surveillance and to help risk assessment studies, with the ultimate objective of protecting consumers and enhancing the microbial safety of bivalve molluscan shellfish as a healthy food.

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A multiplex PCR for the detection of Vibrio vulnificus hazardous to human and/or animal health from seafood

Cited by 12 publications

References 26 publications

Multiplex Analysis of Animal Species Origins in Feedstuffs and Foodstuffs for the Prevention, Control and Eradication of Transmissible Spongiform Encephalopathies (TSEs) by PCR

Multiplex Analysis of Animal Species Origins in Feedstuffs and Foodstuffs for the Prevention, Control and Eradication of Transmissible Spongiform Encephalopathies (TSEs) by PCR

A Novel RAA Combined Test Strip Method Based on Dual Gene Targets for Pathogenic Vibrio vulnificus in Aquatic Products

A Comprehensive Review for the Surveillance of Human Pathogenic Microorganisms in Shellfish

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