title: Development of a real-time loop-mediated isothermal amplification assay for the sensitive and rapid detection of Listeria monocytogenes.
abstract: 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 Listeria 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.
  
  
  CONCLUSIONS
  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 ESEtube 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.
authors: College of Light Industry and Food Sciences, South China University of Technology, Guangzhou, China.
authors: L
authors: L
authors: Ye
authors: College of Light Industry and Food Sciences, South China University of Technology, Guangzhou, China.
authors: Y
authors: Y
authors: Li
authors: College of Environmental and Biological Engineering, Guangdong University of Petrochemical Technology, Maoming, China.
authors: J
authors: J
authors: Zhao
authors: State Key Laboratory of Food Safety Technology for Meat Products, Xiamen Yinxiang Group Co., Ltd., Xiamen, China.
authors: Z
authors: Z
authors: Zhang
authors: College of Light Industry and Food Sciences, South China University of Technology, Guangzhou, China.
authors: H
authors: H
authors: Meng
authors: College of Light Industry and Food Sciences, South China University of Technology, Guangzhou, China.
authors: H
authors: H
authors: Yan
authors: Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University, Tsushima-Naka, Okayama, Japan.
authors: S-I
authors: SI
authors: Miyoshi
authors: College of Light Industry and Food Sciences, South China University of Technology, Guangzhou, China.
authors: L
authors: L
authors: Shi
mesh_terms: D017931:DNA Primers; D005516:Food Microbiology / Q000379:methods*; D006460:Hemolysin Proteins / Q000235:genetics; D006801:Humans; D008089:Listeria monocytogenes / Q000302:isolation & purification*; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D060888:Real-Time Polymerase Chain Reaction
chemical_list: D017931:DNA Primers; D006460:Hemolysin Proteins
keywords: API Listeria; ESEQuant tube scanner; Listeria monocytogenes; RealAmp; real-time PCR