Development of a multiplex real-time PCR method for simultaneous detection of Vibrio parahaemolyticus, Listeria monocytogenes and Salmonella spp. in raw shrimp

Zhang, Zhaohuan; Xiao, Lili; Lou, Yang; Jin, Mengtong; Liao, Chao; Malakar, Pradeep K.; Pan, Yingjie; Yong, Zhao

doi:10.1016/j.foodcont.2014.11.007

Cited by 79 publications

(51 citation statements)

References 35 publications

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“…This extraction method was modified, where the incubation time in lysozyme was increased to 1 h and the incubation time in proteinase K was increased to 2 h (Ye et al 2013;Zhang et al 2015).…”

Section: Dna Extractionmentioning

confidence: 99%

“…Traditional culture-based methods for detecting and enumerating these two pathogens in food products include enrichment, selective plating, and biochemical confirmation and take between 5 and 7 days (Kim et al 2012;Garrido et al 2013;Ma et al 2014;Zhang et al 2015). Real-time PCR is a faster, more sensitive, less labor-intensive quantitative detection method, and has been widely used for detecting and enumerating many pathogens in food (Feng 2007;Willenburg and Divol 2012;Schnetzinger et al 2013;Xiao et al 2015).…”

Section: Introductionmentioning

confidence: 99%

“…Real-time PCR is a faster, more sensitive, less labor-intensive quantitative detection method, and has been widely used for detecting and enumerating many pathogens in food (Feng 2007;Willenburg and Divol 2012;Schnetzinger et al 2013;Xiao et al 2015). For maximum efficiency, multiplex real-time PCR was developed as an automated highthroughput technique for simultaneous and direct detection of more than two pathogens in the same reaction (Kim et al 2012;Garrido et al 2013;Ma et al 2014;Zhang et al 2015). However, one of the limitations of real-time PCR and multiplex real-time PCR methods for the detection of bacteria in food samples is that both viable and dead cells are detected (Kramer et al 2009;Schnetzinger et al 2013;Macé et al 2013;Salam et al 2014).…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Quantifying viable Vibrio parahaemolyticus and Listeria monocytogenes simultaneously in raw shrimp

Zhang

Liu

Lou

et al. 2015

Appl Microbiol Biotechnol

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A novel TaqMan-based multiplex real-time PCR method combined with propidium monoazide (PMA) treatment was firstly developed for the simultaneous quantification of viable Vibrio parahaemolyticus and Listeria monocytogenes in raw shrimp. The optimization of PMA concentration showed that 100 μM was considered optimal to effectively inhibit 10(8) CFU/mL dead cells of both bacteria. The high specificity of this method was confirmed on tests using 96 target and non-target strains. The optimized assay could detect as low as 10(1)-10(2) CFU/g of each strain on the artificially contaminated shrimp, and its amplification efficiencies were up to 100 and 106 % for V. parahaemolyticus and L. monocytogenes, respectively. Furthermore, this assay has been successfully applied to describe the behavior of these two pathogens in raw shrimps stored at 4 °C. In conclusion, this PMA TaqMan-based multiplex real-time PCR technique, where the whole procedure takes less than 5 h, provides an effective and rapid tool for monitoring contamination of viable V. parahaemolyticus and L. monocytogenes in seafood, improving seafood safety and protecting public health.

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Section: Dna Extractionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Quantifying viable Vibrio parahaemolyticus and Listeria monocytogenes simultaneously in raw shrimp

Zhang

Liu

Lou

et al. 2015

Appl Microbiol Biotechnol

Self Cite

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“…The polymerase chain reaction (PCR) assays is one of the molecular techniques that is widely used to detect the presences of pathogenic V. parahaemolyticus strain in food and environment (Panicker et al, 2004;Yamamoto et al, 2008;Paydar et al, 2013;Malcolm et al, 2015). PCR primers can be multiplexed in a single reaction to increase the detection limit or tailored as real-time PCR analysis to provide more rapid results (Grant et al, 2006;Zhang et al, 2014).…”

Section: Introductionmentioning

confidence: 99%

Antimicrobial resistance profile and resistance genes of Vibrio species isolated from giant freshwater prawn (Macrobrachium rosenbergii) raised in Iran

Shakerian

Barton

Akinbowale

et al. 2018

J Hellenic Vet Med Soc

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Because of raising of large-scale high density prawn aquaculture techniques, production of this prawn worldwide is facing a serious threat from fatal diseases caused by nodaviruses and bacteria, particularly from the Vibrio species. The development of antibiotic resistance by Vibrio represents a potential threat to human health by exchange of resistant genes to human pathogens through food chain. This study aimed to determine antibiotic resistance profile of Vibrio isolates from giant fresh water Prawns (Macrobrachium rosenbergii) raised in Iran and to detect some antibiotic resistance genes in the isolates. A total of fifty giant fresh water prawns were processed for isolation of Vibrio species during February 2015 to August 2015. Identification of Vibrio isolates was done following standard biochemical methods. Phenotypic resistance of the isolates as determined by agar dilution method while polymerase chain reaction (PCR) method was used to detect the presence of erm, tetS, strA and sul2 genes in the isolates. Out of 50 prawns, 31 (62%) isolates of Vibrio spp. were reported, of which 20 (40%) were identified as V. parahaemolyticus, 10 (20%) were V. vulnificus and 1 (2%) were V. cholera. Over 90% of the tested strains showed susceptibility to SXT, AZM and NIT. In addition, strA and tetS genes were detected in all isolated Vibrio species. StrA gene was identified in 6 V. parahaemolyticus strains and also ermB and sul2 genes were not present in the isolate of V. cholera. The occurrence of multidrug resistance strains in the environment could be an indication of excessive usage of antibiotics in agriculture and aquaculture fields. This study has shown that giant freshwater prawns raised in Iran habour multidrug resistant Vibrio species.

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“…[5][6][7] Although conventional culturing ensures the accuracy of determinations, and are optimum methods for VP identification, there still exist some shortcomings, such as long-time consumption and cumbersome practical application. Various approaches have been developed to obtain better performances of VP analysis, including enzyme-linked immunesorbent assay (ELISA), [8][9][10] DNA probe, 11 most probable number (MPN), 12,13 polymerase chain reaction (PCR), [14][15][16][17][18] loop-mediated isothermal amplification (LAMP), [19][20][21][22][23] and electrochemistry (EC). 24,25 Despite each of these new approaches having a combination of excellent sensitivity, accuracy and specificity, they still suffer drawbacks involving high analytical cost, need for expensive equipment, and professional trained personnel.…”

Section: Introductionmentioning

confidence: 99%

Christmas-tree Derived Amplification Immuno-Strategy for Sensitive Visual Detection of Vibrio parahaemolyticus Based on Gold Label Silver Stain Technology

Song

et al. 2017

ANAL. SCI.

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Recently, Liu and his coworkers designed a Fe(III)-grafted g-C3N4 photocatalyst that displayed an excellent photocatalytic performance. 37 These make it promising in the development of biosensors with high 2017 © The Japan Society for Analytical Chemistry † To whom correspondence should be addressed. A developed Christmas-tree derived immunosensor based on a gold label silver stain (GLSS) technique was fabricated for a highly sensitive analysis of Vibrio parahaemolyticu (VP). In this strategy, captured VP antibody (cAb) was immobilized on a solid substrate; then, the VPs were sequentially tagged with a signal probe by incubating the assay with a detection VP antibody (dAb) conjugated gold nanoparticles (AuNPs)-labeled graphite-like carbon nitride (g-C3N4). Finally, the attached signal probe could harvest a visible signal by the silver meal deposition, and then followed by homebrew Matlab 6.0 as a grey value acquisition. In addition, the overall design of the biosensor was established in abundant AuNPs and g-C3N4 with a two-dimensional structure, affording a bulb-decorated Christmas-tree model. Moreover, with the optimized conditions, the detection limit of the as-proposed biosensor is as low as 10 2 CFU (Colony-Forming Units) mL -1 , exhibiting an increase of two orders of magnitude compared with the traditional immune-gold method. Additionally, the developed visible immunosensor was also successfully applied to the analysis of complicated samples.

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Development of a multiplex real-time PCR method for simultaneous detection of Vibrio parahaemolyticus, Listeria monocytogenes and Salmonella spp. in raw shrimp

Cited by 79 publications

References 35 publications

Quantifying viable Vibrio parahaemolyticus and Listeria monocytogenes simultaneously in raw shrimp

Quantifying viable Vibrio parahaemolyticus and Listeria monocytogenes simultaneously in raw shrimp

Antimicrobial resistance profile and resistance genes of Vibrio species isolated from giant freshwater prawn (Macrobrachium rosenbergii) raised in Iran

Christmas-tree Derived Amplification Immuno-Strategy for Sensitive Visual Detection of Vibrio parahaemolyticus Based on Gold Label Silver Stain Technology

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