Publication:
Rapid and sensitive detection of Vibrio cholerae by loop-mediated isothermal amplification targeted to the gene of outer membrane protein ompW

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Issued Date

2010

Resource Type

Article

File Type

application/pdf

ISSN

2668254

DOI

10.1111/j.1472-765X.2009.02749.x

Other identifier(s)

2-s2.0-72149112769

Rights Holder(s)

Scopus

Bibliographic Citation

Letters in Applied Microbiology. Vol 50, No.1 (2010), p.36-42

Suggested Citation

Srisuk C., Chaivisuthangkura P., Rukpratanporn S., Longyant S., Sridulyakul P., Sithigorngul P. Rapid and sensitive detection of Vibrio cholerae by loop-mediated isothermal amplification targeted to the gene of outer membrane protein ompW. Letters in Applied Microbiology. Vol 50, No.1 (2010), p.36-42. doi:10.1111/j.1472-765X.2009.02749.x Retrieved from: https://hdl.handle.net/20.500.14740/7661

Title

Rapid and sensitive detection of Vibrio cholerae by loop-mediated isothermal amplification targeted to the gene of outer membrane protein ompW

Author(s)

Srisuk C.
 Chaivisuthangkura P.
 Rukpratanporn S.
 Longyant S.
 Sridulyakul P.
 Sithigorngul P.

Abstract

Aims: The present study was aimed to develop a loop-mediated isothermal amplification (LAMP) assay for rapid and specific detection of Vibrio cholerae. Methods and Results: A set of five designed primers that recognized specifically the V. cholerae ompW gene was used. The optimized time and temperature conditions for the LAMP assay were 75 min at 65°C, respectively. The LAMP method accurately identified 16 isolates of V. cholerae but did not detect 28 non-cholerae Vibrio isolates and 37 non-Vibrio bacterial isolates. The sensitivity of LAMP for V. cholerae detection in pure cultures was 2.2 × 103 CFU ml-1 or equivalent to 8 CFU per reaction. In the case of spiked shrimp samples without enrichment, the detection limit for V. cholerae was 2.2 × 104 CFU g-1 or equivalent to 20 CFU per reaction, while that of PCR was 100 CFU per reaction. Conclusion: The developed LAMP assay targeting ompW gene was rapid, specific and sensitive for V. cholerae detection. Significant and Impact of the study: The developed LAMP assay appears to be precise, accurate and a valuable tool for detection of V. cholerae. This assay can replace laborious biochemical tests for the identification of V. cholerae in contaminated food sample. © 2009 The Society for Applied Microbiology.

Subject(s)

Bacterial isolates
 Biochemical tests
 Detection limits
 Food samples
 Loop-mediated isothermal amplification
 Outer membrane protein
 PCR
 Pure culture
 Sensitive detection
 Specific detection
 Temperature conditions
 Vibrio cholerae
 Amplification
 Ompw protein
 Outer membrane protein
 Unclassified drug
 Amplification
 Bacterium
 Bioassay
 Gene
 Polymerase chain reaction
 Protein
 Sampling
 Article
 Bacterial gene
 Bacterium culture
 Bacterium detection
 Chemical reaction
 Colony forming unit
 Controlled study
 Diagnostic accuracy
 Enrichment culture
 Gene amplification
 Gene sequence
 Genetic analysis
 High temperature
 Listonella anguillarum
 Loop mediated isothermal amplification
 Microbiological examination
 Nonhuman
 Nucleotide sequence
 Sensitivity and specificity
 Vibrio
 Vibrio alginolyticus
 Vibrio campbellii
 Vibrio cholerae
 Vibrio fluvialis
 Vibrio harveyi
 Vibrio mimicus
 Vibrio ordalii
 Vibrio parahaemolyticus
 Vibrio shilonii
 Vibrio vulnificus
 Animals
 Bacterial Outer Membrane Proteins
 Bacterial Typing Techniques
 DNA, Bacterial
 Food Microbiology
 Genes, Bacterial
 Limit of Detection
 Nucleic Acid Amplification Techniques
 Penaeidae
 Polymerase Chain Reaction
 Sensitivity and Specificity
 Shellfish
 Vibrio
 Vibrio cholerae
 Bacteria (microorganisms)
 Decapoda (Crustacea)
 Vibrio
 Vibrio cholerae

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URI

https://hdl.handle.net/20.500.14740/7661

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Scopus: Year 1983-2021

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