Publication:
Rapid and sensitive detection of Vibrio vulnificus by loop-mediated isothermal amplification combined with lateral flow dipstick targeted to rpoS gene

0
0

Issued Date

2011

Resource Type

Article

File Type

application/pdf

ISSN

8908508

DOI

10.1016/j.mcp.2011.04.001

Other identifier(s)

2-s2.0-79959774063

Rights Holder(s)

มหาวิทยาลัยศรีนครินทรวิโรฒ

Bibliographic Citation

Molecular and Cellular Probes. Vol 25, No.4 (2011), p.158-163

Suggested Citation

Surasilp T., Longyant S., Rukpratanporn S., Sridulyakul P., Sithigorngul P., Chaivisuthangkura P. Rapid and sensitive detection of Vibrio vulnificus by loop-mediated isothermal amplification combined with lateral flow dipstick targeted to rpoS gene. Molecular and Cellular Probes. Vol 25, No.4 (2011), p.158-163. doi:10.1016/j.mcp.2011.04.001 Retrieved from: https://hdl.handle.net/20.500.14740/7279

Title

Rapid and sensitive detection of Vibrio vulnificus by loop-mediated isothermal amplification combined with lateral flow dipstick targeted to rpoS gene

Author(s)

Surasilp T.
 Longyant S.
 Rukpratanporn S.
 Sridulyakul P.
 Sithigorngul P.
 Chaivisuthangkura P.

Abstract

A novel loop-mediated isothermal amplification (LAMP) combined with amplicon detection by chromatographic lateral flow dipstick (LFD) assay was developed and evaluated for the detection of Vibrio vulnificus. Biotinylated LAMP amplicons were produced by a set of six designed primers that recognized the V. vulnificus RNA polymerase subunit sigma factor S (rpoS) gene followed by hybridization with an FITC-labeled probe and LFD detection. The optimized time and temperature conditions for the LAMP assay were 90 min at 65 °C. The LAMP-LFD method accurately identified 14 isolates of V. vulnificus but did not detect 25 non-vulnificus Vibrio isolates and 37 non-Vibrio isolates. The sensitivity of LAMP-LFD for V. vulnificus detection in pure culture was 1.5 × 103 CFU ml-1 or equivalent to 2.8 CFU per reaction. In the case of spiked oyster samples without enrichment, the detection limit for V. vulnificus was 1.2 × 104 CFU g-1 or equivalent to 11 CFU per reaction. The results show that this method appears to be accurate, precise and valuable tool for identification of V. vulnificus and can be used efficiently for detection of V. vulnificus in contaminated food sample. © 2011 Elsevier Ltd.

Subject(s)

Bacterial DNA
 Fluorescein isothiocyanate
 RNA polymerase
 RNA polymerase subunit sigma factor s
 Unclassified drug
 Amplicon
 Article
 Bacterial gene
 Bacterium culture
 Bacterium detection
 Bacterium isolate
 Gene targeting
 Listonella anguillarum
 Loop mediated isothermal amplification
 Nonhuman
 Oyster
 Priority journal
 RNA hybridization
 Temperature
 Vibrio
 Vibrio alginolyticus
 Vibrio campbellii
 Vibrio cholerae
 Vibrio fluvialis
 Vibrio harveyi
 Vibrio mimicus
 Vibrio ordalii
 Vibrio parahaemolyticus
 Vibrio shilonii
 Vibrio vulnificus
 Bacterial Proteins
 DNA Primers
 Fluorescein-5-isothiocyanate
 Molecular Probes
 Molecular Sequence Data
 Nucleic Acid Amplification Techniques
 Sigma Factor
 Temperature
 Vibrio vulnificus
 Ostreidae
 Vibrio
 Vibrio vulnificus

View online Resources

Research Projects

Organizational Units

Journal Issue

URI

https://hdl.handle.net/20.500.14740/7279

Collections

Scopus: Year 1983-2021

Endorsement

Review

Supplemented By

Referenced By