Publication: The development of loop-mediated isothermal amplification combined with lateral flow dipstick for detection of Vibrio parahaemolyticus
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Issued Date
2011
Resource Type
File Type
application/pdf
ISSN
2668254
Other identifier(s)
2-s2.0-79952507510
Rights Holder(s)
มหาวิทยาลัยศรีนครินทรวิโรฒ
Bibliographic Citation
Letters in Applied Microbiology. Vol 52, No.4 (2011), p.344-351
Suggested Citation
Prompamorn P., Sithigorngul P., Rukpratanporn S., Longyant S., Sridulyakul P., Chaivisuthangkura P. The development of loop-mediated isothermal amplification combined with lateral flow dipstick for detection of Vibrio parahaemolyticus. Letters in Applied Microbiology. Vol 52, No.4 (2011), p.344-351. doi:10.1111/j.1472-765X.2011.03007.x Retrieved from: https://hdl.handle.net/20.500.14740/7347
Abstract
Aims: The current study was aimed to develop a loop-mediated isothermal amplification (LAMP) combined with amplicon detection by chromatographic lateral flow dipstick (LFD) assay for rapid and specific detection of Vibrio parahaemolyticus. Methods and Results: Biotinylated LAMP amplicons were produced by a set of four designed primers that recognized specifically the V. parahaemolyticus thermolabile haemolysin (tlh) gene followed by hybridization with an FITC-labelled probe and LFD detection. The optimized time and temperature conditions for the LAMP assay were 90min at 65°C. The LAMP-LFD method accurately identified 28 isolates of V. parahaemolyticus but did not detect 24 non-parahaemolyticus Vibrio isolates and 35 non-Vibrio bacterial isolates. The sensitivity of LAMP-LFD for V. parahaemolyticus detection in pure cultures was 120CFUml-1. In the case of spiked shrimp samples without enrichment, the detection limit for V. parahaemolyticus was 1·8×103CFUg-1 or equivalent to 3CFU per reaction while that of conventional PCR was 30CFU per reaction. Conclusions: The established LAMP-LFD assay targeting tlh gene was specific, rapid and sensitive for identification of V. parahaemolyticus. Significance and Impact of the Study: The developed LAMP-LFD assay provided a valuable tool for detection of V. parahaemolyticus and can be used effectively for identification of V. parahaemolyticus in contaminated food sample. © 2011 The Authors. Letters in Applied Microbiology © 2011 The Society for Applied Microbiology.
Subject(s)
Bacterium
Gene
Hybridization
Optimization
Polymerase chain reaction
Amplicon
Article
Assay
Chromatographic lateral flow dipstick assay
Colony forming unit
Food contamination
Gene
Loop mediated isothermal amplification
Nonhuman
Sensitivity analysis
Thermolabile hemolysin gene
Vibrio parahaemolyticus
DNA Primers
Hemolysin Proteins
Nucleic Acid Amplification Techniques
Nucleic Acid Hybridization
Polymerase Chain Reaction
Temperature
Vibrio parahaemolyticus
Bacteria (microorganisms)
Decapoda (Crustacea)
Vibrio
Vibrio parahaemolyticus
Gene
Hybridization
Optimization
Polymerase chain reaction
Amplicon
Article
Assay
Chromatographic lateral flow dipstick assay
Colony forming unit
Food contamination
Gene
Loop mediated isothermal amplification
Nonhuman
Sensitivity analysis
Thermolabile hemolysin gene
Vibrio parahaemolyticus
DNA Primers
Hemolysin Proteins
Nucleic Acid Amplification Techniques
Nucleic Acid Hybridization
Polymerase Chain Reaction
Temperature
Vibrio parahaemolyticus
Bacteria (microorganisms)
Decapoda (Crustacea)
Vibrio
Vibrio parahaemolyticus