A single fluorescence-based LAMP reaction for identifying multiple parasites in mosquitoes

Vector-borne diseases, such as malaria and lymphatic filariasis, are co-endemic in large parts of the world. To develop a multiplex amplification method for the simultaneous detection of multiple insect-borne infectious diseases, we used LAMP with fluorescently labeled primers to identify the SPECT2 gene of Plasmodium berghei and the cytochrome oxidase subunit I gene of Dirofilaria immitis in mosquitoes. This technique could detect as few as 100 P. berghei-infected red blood cell-equivalents or one D. immitis microfilaria. Moreover, individual species of parasites in mosquitoes could be identified when a mixture of fluorescently labeled primer sets was used. These findings suggest that the multiplex LAMP assay is sensitive and specific enough to identify parasite-bearing mosquitoes in areas where several diseases occur simultaneously. This procedure could increase the efficiency and effectiveness of arthropod-borne disease elimination programs. (C) 2010 Elsevier Inc. All rights reserved.

媒介传播疾病，如疟疾和淋巴丝虫病，在世界大部分地区共同流行。为了开发一种多重扩增方法用于同时检测多种虫媒传染病，我们使用带有荧光标记引物的环介导等温扩增技术（LAMP）来识别伯氏疟原虫的SPECT2基因以及犬恶丝虫的细胞色素氧化酶亚基I基因。这种技术能够检测到少至100个相当于伯氏疟原虫感染的红细胞或1条犬恶丝虫微丝蚴。此外，当使用混合的荧光标记引物组时，可以识别蚊子体内的单个寄生虫种类。这些发现表明，多重LAMP检测具有足够的敏感性和特异性，能够在多种疾病同时发生的地区识别携带寄生虫的蚊子。这一程序可以提高虫媒疾病消除计划的效率和效果。（C）2010爱思唯尔公司。保留所有权利。