开发一种用于灵敏快速检测单核细胞增生李斯特菌的实时环介导等温扩增检测方法。
Development of a real-time loop-mediated isothermal amplification assay for the sensitive and rapid detection of Listeria monocytogenes.
作者信息
Ye L, Li Y, Zhao J, Zhang Z, Meng H, Yan H, Miyoshi S-I, Shi L
机构信息
College of Light Industry and Food Sciences, South China University of Technology, Guangzhou, China.
College of Environmental and Biological Engineering, Guangdong University of Petrochemical Technology, Maoming, China.
出版信息
Lett Appl Microbiol. 2015 Jul;61(1):85-90. doi: 10.1111/lam.12429. Epub 2015 May 10.
UNLABELLED
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.
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 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.
未加标签
开发了一种用于检测李斯特菌的实时环介导等温扩增（RealAmp）检测方法。使用针对编码溶血素的hlyA基因的特异性引物的RealAmp检测方法，用来自世界不同地区的58株单核细胞增生李斯特菌菌株进行了验证。RealAmp检测方法的灵敏度和特异性都很高。RealAmp检测方法能够在30分钟内检测到10(3) CFU ml(-1)。对RealAmp检测方法、API李斯特菌检测方法和实时PCR检测方法的比较评估表明，RealAmp检测方法比其他两种检测方法更简单、更快，且具有更高的特异性。
研究的意义和影响
传统的培养和分子检测方法总是耗时且需要特定的实验室基础设施，从而限制了它们用于病原体的快速检测和诊断。开发了一种通过ESEtube扫描仪进行的实时环介导等温扩增（RealAmp）检测方法，以快速检测从食品中分离出的单核细胞增生李斯特菌。结果表明，使用试管扫描仪的RealAmp检测方法比传统的API李斯特菌检测方法和实时PCR检测方法更高效、精确。
Zotero 插件