Division of Bacteriology & Mycology, Indian Veterinary Research Institute, Izatnagar, UP, India.
Division of Veterinary Biotechnology, Indian Veterinary Research Institute, Izatnagar, UP, India.
J Appl Microbiol. 2019 May;126(5):1332-1339. doi: 10.1111/jam.13938. Epub 2019 Mar 20.
To develop a rapid, sensitive and specific diagnostic assay for the detection of Brucella.
The probe-based RT-LAMP was carried out by using a set of four or six primers and different LAMP chemicals to compare its results with real-time PCR. Detection of gene amplification is done within 40 min and can be seen by amplification curve, turbidity and addition of DNA-binding dye at the end of the reaction results in colour difference under normal day light and in UV. The sensitivity of probe-based real-time LAMP assay was found 10-fold higher than Taqman-based qPCR. The specificity of the developed assay was validated by the absence of any cross-reaction with other pathogenic bacteria.
The developed probe-based RT-LAMP assay is extremely rapid, cost effective, highly specific and sensitive, and has potential usefulness for rapid Brucella surveillance.
The developed probe-based RT-LAMP is a powerful gene amplification technique which is a specific, fast diagnostic tool for early detection and identification of Brucella.
开发一种用于检测布鲁氏菌的快速、灵敏和特异的诊断检测方法。
采用一组四或六对引物和不同的 LAMP 化学试剂进行基于探针的 RT-LAMP，并将其结果与实时 PCR 进行比较。基因扩增的检测在 40 分钟内完成，可以通过扩增曲线、浊度和在反应结束时添加 DNA 结合染料来观察，在正常日光和紫外线下会产生颜色差异。基于探针的实时 LAMP 检测方法的灵敏度比 Taqman 基 qPCR 高 10 倍。通过与其他致病性细菌无交叉反应验证了所开发检测方法的特异性。
所开发的基于探针的 RT-LAMP 检测方法快速、具有成本效益、高度特异和灵敏，对于快速进行布鲁氏菌监测具有潜在的用途。
所开发的基于探针的 RT-LAMP 是一种强大的基因扩增技术，是一种用于早期检测和鉴定布鲁氏菌的特异性、快速的诊断工具。
