Center of Excellence for Shrimp Molecular Biology and Biotechnology (CENTEX Shrimp), Faculty of Science, Mahidol University, Bangkok, Thailand; National Center for Genetic Engineering and Biotechnology (BIOTEC), National Science and Technology Development Agency, Pathum Thani, Thailand.
Lett Appl Microbiol. 2013 Oct;57(4):317-24. doi: 10.1111/lam.12114. Epub 2013 Jun 26.
Streptococcus agalactiae and Strep. iniae are bacterial pathogens that cause streptococcosis in many fish species. An accelerated colorimetric loop-mediated isothermal amplification (LAMP) assay with pre-addition of calcein was established, and the transmission and detection of Strep. agalactiae and Strep. iniae in tilapia under natural aquatic environment were investigated. A positive reaction was observed by a colour change from orange to green through the naked eyes after completion at 63°C for 30 min with 10 times higher sensitivity than that of nested PCR assays and without cross-amplification with other fish bacterial pathogens. All sample types of Nile and red tilapia (broodstock, fertilized egg, fry) were Strep. agalactiae- and Strep. iniae positive by this new method, implying that they could be vertically transmitted. With its application for screening broodstock and fry before stocking and for monitoring fish health in grow-out ponds, the method would become very useful in fish farming industry.
The application of colorimetric LAMP with pre-addition of calcein offers simple, rapid and sensitive technique with applicability for small field laboratories. This technique explored the possible vertical transmission mode of Strep. agalactiae and Strep. iniae under natural aquatic environment. It could be such preliminary data provided for the screening broodstock before breeding and/or the specific-pathogen-free production.
无乳链球菌和酿脓链球菌是细菌性病原体，可导致许多鱼类发生链球菌病。建立了一种加速比色环介导等温扩增（LAMP）测定法，并在自然水生环境下研究了罗非鱼和虾中无乳链球菌和酿脓链球菌的传播和检测。在 63°C 下完成 30 分钟后，通过肉眼观察到颜色从橙色变为绿色，发生阳性反应，比巢式 PCR 检测方法高 10 倍，与其他鱼类细菌病原体无交叉扩增。新方法检测尼罗罗非鱼和红罗非鱼（亲鱼、受精卵、鱼苗）的所有样本类型均为无乳链球菌和酿脓链球菌阳性，这表明它们可能垂直传播。该方法可用于在放养前筛选亲鱼和鱼苗，并监测养殖池塘中的鱼类健康，因此在水产养殖行业非常有用。
添加钙黄绿素的比色 LAMP 的应用提供了一种简单、快速和敏感的技术，适用于小型现场实验室。该技术在自然水生环境下探索了无乳链球菌和酿脓链球菌可能的垂直传播模式。它可以为繁殖前筛选亲鱼和/或无特定病原体生产提供初步数据。
