Publication: Rapid and sensitive detection of Vibrio alginolyticus by loop-mediated isothermal amplification combined with a lateral flow dipstick targeted to the rpoX gene
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Issued Date
2015
Resource Type
File Type
application/pdf
ISSN
8997659
Other identifier(s)
2-s2.0-84940045331
Rights Holder(s)
Scopus
Bibliographic Citation
Journal of Aquatic Animal Health. Vol 27, No.3 (2015), p.156-163
Suggested Citation
Plaon S., Longyant S., Sithigorngul P., Chaivisuthangkura P. Rapid and sensitive detection of Vibrio alginolyticus by loop-mediated isothermal amplification combined with a lateral flow dipstick targeted to the rpoX gene. Journal of Aquatic Animal Health. Vol 27, No.3 (2015), p.156-163. doi:10.1080/08997659.2015.1037468 Retrieved from: https://hdl.handle.net/20.500.14740/6250
Abstract
Vibrio alginolyticus is a major bacterial pathogen causing disease in marine animals. The present study aimed to develop a loop-mediated isothermal amplification (LAMP) coupled with a lateral flow dipstick (LFD) for rapid and simple visual detection of V. alginolyticus–specific amplicons. The biotin-labeled LAMP amplicons from the targeted portion of a gene encoding rpoS-like sigma factor (rpoX) were generated at 60°C for 1 h and then hybridized with a fluorescein isothiocyanate–labeled probe for 5 min for visual detection with LFD. In pure cultures, the detection limit of the LAMP–LFD technique for V. alginolyticus was 1.8 × 102 CFU/mL while that of PCR was 1.8 × 103 CFU/mL. In spiked whiteleg shrimp samples Penaeus vannamei, the sensitivity for V. alginolyticus detection was 2 × 103 CFU/g (equivalent to 4 CFU per reaction) while PCR was 10 times less sensitive. The LAMP–LFD method for V. alginolyticus correctly identified 21 isolates of V. alginolyticus but did not recognize 23 non-V. alginolyticus Vibrio isolates and 15 non-Vibrio isolates. In summary, this LAMP–LFD method targeted to the rpoX gene is a convenient assay for specific identification of V. alginolyticus with high sensitivity. © American Fisheries Society 2015.
Subject(s)
Animalia
Bacteria (microorganisms)
Litopenaeus vannamei
Vibrio
Vibrio alginolyticus
Bacterial protein
Molecular probe
Chemistry
Gene expression regulation
Genetics
Isolation and purification
Metabolism
Molecular probe
Nucleic acid amplification
Physiology
Procedures
Vibrio alginolyticus
Bacterial Proteins
Gene Expression Regulation, Bacterial
Molecular Probes
Nucleic Acid Amplification Techniques
Vibrio alginolyticus
Bacteria (microorganisms)
Litopenaeus vannamei
Vibrio
Vibrio alginolyticus
Bacterial protein
Molecular probe
Chemistry
Gene expression regulation
Genetics
Isolation and purification
Metabolism
Molecular probe
Nucleic acid amplification
Physiology
Procedures
Vibrio alginolyticus
Bacterial Proteins
Gene Expression Regulation, Bacterial
Molecular Probes
Nucleic Acid Amplification Techniques
Vibrio alginolyticus