Detection of enteropathogenic Vibrio parahaemolyticus, Vibrio cholerae and Vibrio vulnificus: performance of real-time PCR kits in an interlaboratory study

Eschbach, Erik; Martin, Annett; Huhn, Jennifer; Seidel, Constanze; Heuer, Ralf; Schumacher, Jan-Hendrik; Ulrich, Stefan; Axe, Jens-Oliver; Konietzny, Antje; Strauch, Eckhard; Oberheitmann, Boris

doi:10.1007/s00217-017-2844-z

Cited by 12 publications

(3 citation statements)

References 26 publications

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“…It is a member of the genus Vibrio from family Vibrionaceae , a Gram-negative, marine halophilic bacterium that naturally inhabits in global coastal waters, sediment and various types of marine animals [ 3 ] such as fish, shrimp, crab, clam, oyster [ 4 , 5 , 6 , 7 ], and mussel [ 8 ]. V. parahaemolyticus can also be transmitted to humans when consuming contaminated raw or poorly cooked seafood products [ 9 , 10 ]. This bacterium contributes to an acute gastroenteritis, which includes nausea, diarrhea, vomiting, fever, and chills.…”

Section: Introductionmentioning

confidence: 99%

Vibrio parahaemolyticus Isolates from Asian Green Mussel: Molecular Characteristics, Virulence and Their Inhibition by Chitooligosaccharide-Tea Polyphenol Conjugates

Palamae

Mittal

Yingkajorn

et al. 2022

Foods

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Fifty isolates of Vibrio parahaemolyticus were tested for pathogenicity, biofilm formation, motility, and antibiotic resistance. Antimicrobial activity of chitooligosaccharide (COS)-tea polyphenol conjugates against all isolates was also studied. Forty-three isolates were randomly selected from 520 isolates from Asian green mussel (Perna viridis) grown on CHROMagarTM Vibrio agar plate. Six isolates were acquired from stool specimens of diarrhea patients. One laboratory strain was V. parahaemolyticus PSU.SCB.16S.14. Among all isolates tested, 12% of V. parahaemolyticus carried the tdh+trh– gene and were positive toward Kanagawa phenomenon test. All of V. parahaemolyticus isolates could produce biofilm and showed relatively strong motile ability. When COS-catechin conjugate (COS-CAT) and COS-epigallocatechin-3-gallate conjugate (COS-EGCG) were examined for their inhibitory effect against V. parahaemolyticus, the former showed the higher bactericidal activity with the MBC value of 1.024 mg/mL against both pathogenic and non-pathogenic strains. Most of the representative Asian green mussel V. parahaemolyticus isolates exhibited high sensitivity to all antibiotics, whereas one isolate showed the intermediate resistance to cefuroxime. However, the representative clinical isolates were highly resistant to nine types of antibiotics and had multiple antibiotic resistance (MAR) index of 0.64. Thus, COS-CAT could be used as potential antimicrobial agent for controlling V. parahaemolyticus-causing disease in Asian green mussel.

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Section: Introductionmentioning

confidence: 99%

Vibrio parahaemolyticus Isolates from Asian Green Mussel: Molecular Characteristics, Virulence and Their Inhibition by Chitooligosaccharide-Tea Polyphenol Conjugates

Palamae

Mittal

Yingkajorn

et al. 2022

Foods

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“…Several PCR methods are available for the specific detection of Salmonella spp. [32,33], V. cholerae [34,35], and V. parahaemolyticus [35,36]. However, identification of individual pathogens by PCR becomes time-consuming and costly.…”

Section: Evaluation Of Multiplex Pcr For Detection Of Seafood Pathogensmentioning

confidence: 99%

Rapid detection of Salmonella contamination in seafoods using multiplex PCR

Sahu¹,

Singh

Behera

et al. 2019

Braz J Microbiol

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Effective monitoring of Salmonella contamination in seafood processing to conform the requirements of HACCP is a great challenge today. Such challenges can be effectively addressed, if the conventional detection methods are replaced with DNAbased molecular methods. Accordingly, it was aimed to develop a robust PCR protocol for specific detection of Salmonella spp. Out of the different primers screened, one pair of primers developed in this study targeting invA gene demonstrated 100% inclusivity for a wide range of Salmonella serotypes and 100% exclusivity for wide range of non-target species. The in silico analysis of the nucleotide sequence obtained from the PCR product suggests its potential as a hybridization probe for genus specific detection of Salmonella spp. contamination. The PCR protocol was sensitive enough to detect 15 cells per reaction using crude DNA prepared within a short time directly from artificially contaminated shrimp tissue. The study demonstrated that the result of PCR reaction can come out on the same day of sample arrival. Incorporation of this pair of primers in a multiplex PCR designed for simultaneous detection of four common seafood-borne human pathogens yielded 147 bp, 302 bp, 403 bp, and 450 bp distinct DNA bands specifically targeting E. coli, toxigenic Vibrio cholerae, Salmonella spp., and V. parahaemolyticus, respectively in a single PCR tube. The PCR methods developed in this study has the potential to be used in the seafood processing plants for effective monitoring of CCPs required for implementation of HACCP-based quality assurance system.

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“…This bacterium is widely distributed in the marine and estuarine environment ( Bej et al, 1999 ) and is a major food borne pathogen in China and South East Asia countries ( Su and Liu, 2007 ; Zhang et al, 2015a ). It is the common cause of gastrointestinal disease from consumption of raw or lightly cooked seafood ( Letchumanan et al, 2015 ; Eschbach et al, 2017 ). The V. parahaemolyticus virulence factors contributing to disease include adhesin, thermostable direct hemolysin (TDH), TDH related hemolysin (TRH), 2 type III secretion systems (T3SS1 and T3SS2) and 2 type VI secretion systems (T6SS1 and T6SS) ( Ceccarelli et al, 2013 ; Wang R. et al, 2015 ).…”

Section: Introductionmentioning

confidence: 99%

A Novel qPCR Method for Simultaneous Detection and Quantification of Viable Pathogenic and Non-pathogenic Vibrio parahaemolyticus (tlh+, tdh+, and ureR+)

et al. 2018

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Pathogenic and non-pathogenic Vibrio parahaemolyticus strains were simultaneously detected and quantified using a novel viable multiplex real-time PCR (novel qPCR). We used a new PCR primer and probe, ureR, as a surrogate for detection of the toxin trh gene as the primer was better at identifying variant V. parahaemolyticus trh strains. The specificity of all primers and probes used in this study were validated on three standard strains of V. parahaemolyticus, 42 clinical strains, 12 wild strains, 4 strains of Vibrio spp., and 4 strains of other bacteria. Then, propidium monoazide (PMA) was applied to inhibit DNA of dead cell, and the results of PMA optimized treatments were 15 μM concentration, 5 min incubation periods, 15 min light exposure periods and 30 RPM rotational speed, which resulted in time and cost savings. Pathogenic and non-pathogenic strains were quantified using a two-reaction tube method where the tlh, tdh, and ureR genes were amplified. Additionally, standard curves with a 7-log dynamic range were generated for quantifying viable V. parahaemolyticus and the amplification efficiencies were 108.68, 105.17, and 115.61% for tlh+, tdh+, and ureR+. This novel qPCR accurately monitored V. parahaemolyticus contamination rates in shrimps (Penaeus vannamei) and clams (Ruditapes philippinarum) sampled from retail stores located in a major district in Shanghai. In conclusion, our assay can prioritize the detection and quantification of viable pathogenic V. parahaemolyticus and can prove to be a more effective tool for reducing infection risks from consumption of seafood in Shanghai.

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Detection of enteropathogenic Vibrio parahaemolyticus, Vibrio cholerae and Vibrio vulnificus: performance of real-time PCR kits in an interlaboratory study

Cited by 12 publications

References 26 publications

Vibrio parahaemolyticus Isolates from Asian Green Mussel: Molecular Characteristics, Virulence and Their Inhibition by Chitooligosaccharide-Tea Polyphenol Conjugates

Vibrio parahaemolyticus Isolates from Asian Green Mussel: Molecular Characteristics, Virulence and Their Inhibition by Chitooligosaccharide-Tea Polyphenol Conjugates

Rapid detection of Salmonella contamination in seafoods using multiplex PCR

A Novel qPCR Method for Simultaneous Detection and Quantification of Viable Pathogenic and Non-pathogenic Vibrio parahaemolyticus (tlh+, tdh+, and ureR+)

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