Pest and Environmental Adaptation Research Group, School of BioSciences, The University of Melbourne, Victoria, Australia.
PLoS One. 2019 Nov 20;14(11):e0225321. doi: 10.1371/journal.pone.0225321. eCollection 2019.
With Wolbachia-based arbovirus control programs being scaled and operationalised around the world, cost effective and reliable detection of Wolbachia in field samples and laboratory stocks is essential for quality control. Here we validate a modified loop-mediated isothermal amplification (LAMP) assay for routine scoring of Wolbachia in mosquitoes from laboratory cultures and the field, applicable to any setting. We show that this assay is a rapid and robust method for highly sensitive and specific detection of wAlbB Wolbachia infection within Aedes aegypti under a variety of conditions. We test the quantitative nature of the assay by evaluating pooled mixtures of Wolbachia-infected and uninfected mosquitoes and show that it is capable of estimating infection frequencies, potentially circumventing the need to perform large-scale individual analysis for wAlbB infection status in the course of field monitoring. These results indicate that LAMP assays are useful for routine screening particularly under field conditions away from laboratory facilities.
随着基于沃尔巴克氏体的虫媒病毒控制项目在全球范围内扩大和实施，在现场样本和实验室库存中经济高效且可靠地检测沃尔巴克氏体对于质量控制至关重要。在这里，我们验证了一种改良的环介导等温扩增 (LAMP) 检测法，用于常规评分实验室培养物和野外蚊子中的沃尔巴克氏体，适用于任何环境。我们表明，该检测法是一种快速且强大的方法，可在各种条件下高度灵敏和特异性地检测埃及伊蚊中的 wAlbB 沃尔巴克氏体感染。我们通过评估沃尔巴克氏体感染和未感染蚊子的混合样本来测试该检测法的定量性质，并表明它能够估计感染频率，可能避免了在野外监测过程中对 wAlbB 感染状态进行大规模个体分析的需要。这些结果表明，LAMP 检测法特别在远离实验室设施的野外条件下是一种有用的常规筛选方法。
