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Development of a real‐time loop‐mediated isothermal amplification assay for the sensitive and rapid detection of L isteria monocytogenes
Author(s) -
Ye L.,
Li Y.,
Zhao J.,
Zhang Z.,
Meng H.,
Yan H.,
Miyoshi S.i.,
Shi L.
Publication year - 2015
Publication title -
letters in applied microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.698
H-Index - 110
eISSN - 1472-765X
pISSN - 0266-8254
DOI - 10.1111/lam.12429
Subject(s) - listeria monocytogenes , loop mediated isothermal amplification , microbiology and biotechnology , listeria , isothermal process , biology , loop (graph theory) , bacteria , chemistry , chromatography , physics , genetics , mathematics , dna , combinatorics , thermodynamics
A real‐time loop‐mediated isothermal amplification (RealAmp) assay for the detection of Listeria was developed. The RealAmp assay, using primers specific for the hemolysin‐encoding hlyA gene, was verified using L isteria monocytogenes strains ( n = 58) from different regions of the world. Both the sensitivity and specificity of the RealAmp assay were high. The RealAmp assay could detect 10 3 CFU ml −1 within 30 min. A comparative evaluation of the RealAmp assay, the API Listeria assay, and the real‐time PCR assay revealed that the RealAmp assay is simpler, faster, and has a higher specificity than the other two assays. Significance and Impact of the Study Conventional culture and molecular detection methods are always time consuming and require a specific laboratory infrastructure, thereby restricting their use for the rapid detection and diagnosis of pathogens. A real‐time loop‐mediated isothermal amplification (RealAmp) assay performed by ESE tube scanner to rapidly detect Listeria monocytogenes isolated from food was developed. The results showed that the RealAmp assay using the tube scanner was more efficient and precise than the conventional API Listeria assay and the real‐time PCR assay.