Abstract：
Spread of the Anopheles stephensi mosquito, an invasive malaria vector, threatens to put an additional 126 million persons per year in Africa at risk for malaria. To accelerate the early detection and rapid response to this mosquito species, confirming its presence and geographic extent is critical. However, existing molecular species assays require specialized laboratory equipment, interpretation, and sequencing confirmation. We developed and optimized a colorimetric rapid loop-mediated isothermal amplification assay for molecular An. stephensi species identification. The assay requires only a heat source and reagents and can be used with or without DNA extraction, resulting in positive color change in 30-35 minutes. We validated the assay against existing PCR techniques and found 100% specificity and analytical sensitivity down to 0.0003 nanograms of genomic DNA. The assay can successfully amplify single mosquito legs. Initial testing on samples from Marsabit, Kenya, illustrate its potential as an early vector detection and malaria mitigation tool.
摘要：
史蒂芬氏按蚊的传播，一种侵袭性疟疾媒介，有可能使非洲每年额外的1.26亿人面临疟疾风险。为了加快对这种蚊子的早期发现和快速反应，确认它的存在和地理范围是至关重要的。然而,现有的分子物种测定需要专门的实验室设备，解释,和测序确认。我们开发并优化了分子An的比色快速环介导等温扩增测定法。斯蒂芬斯物种鉴定。该测定仅需要热源和试剂，可以在有或没有DNA提取的情况下使用，在30-35分钟内产生正颜色变化。我们针对现有的PCR技术验证了该测定，发现100％的特异性和分析灵敏度低至0.0003纳克的基因组DNA。该测定可以成功地扩增单个蚊子腿。对Marsabit样本的初步测试,肯尼亚,说明其作为早期病媒检测和疟疾缓解工具的潜力。
