期刊
MOLECULAR AND CELLULAR PROBES
卷 25, 期 4, 页码 158-163出版社
ACADEMIC PRESS LTD- ELSEVIER SCIENCE LTD
DOI: 10.1016/j.mcp.2011.04.001
关键词
rpoS gene; Vibrio vulnificus; Loop-mediated isothermal amplification (LAMP); PCR
类别
资金
- Office of Commission on Higher Education, Thailand
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 degrees 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 x 10(3) 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 x 10(4) 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. (C) 2011 Elsevier Ltd. All rights reserved.
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