Evaluation of Most-Probable-Number–PCR Method with Internal Amplification Control for the Counting of Total and Pathogenic Vibrio parahaemolyticus in Frozen Shrimps

Copin, S.; Robert-Pillot, Annick; Malle, Pierre; Quilici, Marie‐Laure

doi:10.4315/0362-028x.jfp-11-165

Cited by 16 publications

(9 citation statements)

References 9 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The most probable number coupled with polymerase chain reaction (MPN-PCR) method detailed in the literature was adapted for determining the total Vibrio spp. density (TVD) in water samples ( Ahmad et al, 2011 ; Copin et al, 2012 ; Ramos et al, 2014 ; Abioye and Okoh, 2018 ). As a statistical-based method, MPN only estimates the viable numbers of bacteria in a sample by the principle of extinction dilution, i.e., inoculating liquid medium in 10-fold dilutions to a dilution factor at which no turbidity is observed after the incubation period.…”

Section: Methodsmentioning

confidence: 99%

Molecular Detection and Distribution of Six Medically Important Vibrio spp. in Selected Freshwater and Brackish Water Resources in Eastern Cape Province, South Africa

2021

View full text Add to dashboard Cite

Water resources contaminated with pathogenic Vibrio species are usually a source of devastating infection outbreaks that have been a public health concern in both developed and developing countries over the decades. The present study assessed the prevalence of six medically significant Vibrio species in some water resources in Eastern Cape Province, South Africa for 12 months. We detected vibrios in all the 194 water samples analyzed using polymerase chain reaction (PCR). The prevalence of Vibrio cholerae, Vibrio mimicus, Vibrio fluvialis, Vibrio vulnificus, Vibrio alginolyticus, and Vibrio parahaemolyticus in freshwater samples was 34, 19, 9, 2, 3, and 2%, and that in brackish water samples was 44, 28, 10, 7, 46, and 51%, respectively. The population of the presumptive Vibrio spp. isolated from freshwater (628) and brackish water (342) samples that were confirmed by PCR was 79% (497/628) and 85% (291/342), respectively. Twenty-two percent of the PCR-confirmed Vibrio isolates from freshwater (n = 497) samples and 41% of the PCR-confirmed Vibrio isolates from brackish water samples (n = 291) fall among the Vibrio species of interest. The incidences of V. cholerae, V. mimicus, V. fluvialis, V. vulnificus, V. alginolyticus, and V. parahaemolyticus amidst these Vibrio spp. of interest that were recovered from freshwater samples were 75, 14, 4, 6, 1, and 1%, whereas those from brackish water samples were 24, 7, 3, 3, 47, and 18%, respectively. Our observation during the study suggests pollution as the reason for the unusual isolation of medically important vibrios in winter. Correlation analysis revealed that temperature drives the frequency of isolation, whereas salinity drives the composition of the targeted Vibrio species at our sampling sites. The finding of the study is of public health importance going by the usefulness of the water resources investigated. Although controlling and preventing most of the factors that contribute to the prevalence of medically important bacteria, such as Vibrio species, at the sampling points might be difficult, regular monitoring for creating health risk awareness will go a long way to prevent possible Vibrio-related infection outbreaks at the sampling sites and their immediate environment.

show abstract

Section: Methodsmentioning

confidence: 99%

Molecular Detection and Distribution of Six Medically Important Vibrio spp. in Selected Freshwater and Brackish Water Resources in Eastern Cape Province, South Africa

2021

View full text Add to dashboard Cite

show abstract

“…The classic method of identifying V. parahaemolyticus relies on a variety of biochemical tests, which are laborious and time‐consuming. Modern identification techniques especially based on molecular biology and immunology are developed . Besides, the development of analytical techniques for fatty acid (FA) has rendered use of fatty‐acid based technology to identify a number of organisms practically .…”

Section: Introductionmentioning

confidence: 99%

Fatty acid profiles of Vibrio parahaemolyticus and its changes with environment

Wang

Mou

2013

J. Basic Microbiol.

View full text Add to dashboard Cite

The fatty acid (FA) profiles of 15 strains representing four genera (Vibrio, Pseudomonas, Aeromonas, and Shewanella) and seven species were compared by capillary gas chromatography with flame ionization detection. FA fingerprints of Vibrio parahaemolyticus were established by similarity calculation and principle component analysis. This provided a simple measure for distinguishing V. parahaemolyticus from other bacteria. The similarity scores indicated by correlation coefficient and vector cosine were divided into three regions and suggested that the strains with scores higher than 0.980 probably belonged to V. parahaemolyticus. However, samples with low scores (<0.850) were classified under other genera. Furthermore, the alterations in FA profiles of V. parahaemolyticus in the presence of various environmental pressures were investigated. The production of saturated FA (SFA) increased gradually concomitant with a decreased proportion of unsaturated FA (UFA) with rising temperature. Similarly, the SFA tended to increase at the expense of UFA with prolonged culture time. In addition, V. parahaemolyticus changed its FA profiles to contain increased short-chained FA to resist an acidic environment, whereas alkaline conditions stimulated high production of long-chained FA. Analysis on FA profile is valuable for the physiological study of V. parahaemolyticus and its rapid identification.

show abstract

“…The LoDs of the LAMP assay revealed that LAMP reactions were not influenced by any inhibitor present in the prepared DNA template solutions from seafood and seawater, whereas PCR reactions were somewhat handicapped by inhibitors (Copin et al . ; Karanis et al . ).…”

Section: Resultsmentioning

confidence: 99%

groEL is a suitable genetic marker for detecting Vibrio parahaemolyticus by loop-mediated isothermal amplification assay

Siddique

Jang

Lee

et al. 2017

Letters in Applied Microbiology

View full text Add to dashboard Cite

Significance and Impact of the Study: The loop-mediated isothermal amplification (LAMP) assay using groEL gene (an abundant, highly conserved gene and member of the groESL chaperone gene family) provided rapid, species-specific and highly sensitive method for detecting Vibrio parahaemolyticus, the leading causal agent of seafood-borne diseases worldwide. Moreover, groEL LAMP revealed high efficiency than conventional PCR assay for V. parahaemolyticus using template both from pure culture and artificially contaminated seafood and water, which indicated the applicability in the field and environmental screening purpose for the organism. AbstractA groEL gene-based loop-mediated isothermal amplification (LAMP) assay was developed to detect Vibrio parahaemolyticus in contaminated seafood and water. The assay was optimized and conducted at 63°C for 40 min using Bacillus stearothermophilus (Bst) DNA polymerase, large fragment. Amplification was analysed via multiple detection methods, including opacity, formation of white precipitate, DNA intercalating dyes (ethidium bromide and SYBR Green I), metal ion-binding indicator dye, calcein, and 2% agarose gel electrophoresis. A characteristic ladder-like band pattern on agarose gel and the desired colour changes when using different dyes were observed in positive cases, and these were species-specific for V. parahaemolyticus when compared with other closely related Vibrio spp. The limit of detection (LoD) of this assay was 100 fg per reaction, 100-fold higher than that for conventional polymerase chain reaction (PCR). When tested on artificially contaminated seafood and seawater, the LoDs of the LAMP assay were 120 and 150 fg per reaction respectively, and those of conventional PCR were 120 and 150 pg per reaction respectively. Based on our results, the groEL gene-based LAMP assay is rapid, specific, sensitive, and reliable for detecting V. parahaemolyticus, and it could be used in field diagnosis.

show abstract

Evaluation of Most-Probable-Number–PCR Method with Internal Amplification Control for the Counting of Total and Pathogenic Vibrio parahaemolyticus in Frozen Shrimps

Cited by 16 publications

References 9 publications

Molecular Detection and Distribution of Six Medically Important Vibrio spp. in Selected Freshwater and Brackish Water Resources in Eastern Cape Province, South Africa

Molecular Detection and Distribution of Six Medically Important Vibrio spp. in Selected Freshwater and Brackish Water Resources in Eastern Cape Province, South Africa

Fatty acid profiles of Vibrio parahaemolyticus and its changes with environment

groEL is a suitable genetic marker for detecting Vibrio parahaemolyticus by loop-mediated isothermal amplification assay

Contact Info

Product

Resources

About