The Vector Population Monitoring Tool (VPMT): High-Throughput DNA-Based Diagnostics for the Monitoring of Mosquito Vector Populations

Abstract

Regular monitoring of mosquito vector populations is an integral component of most vector control programmes. Contemporary data on mosquito species composition, infection status, and resistance to insecticides are a prerequisite for effective intervention. For this purpose we, with funding from the Innovative Vector Control Consortium (IVCC), have developed a suite of high-throughput assays based on a single "closed-tube" platform that collectively comprise the "Vector Population Monitoring Tool" (VPMT). The VPMT can be used to screen mosquito disease vector populations for a number of traits including Anopheles gambiae s.l. and Anopheles funestus species identification, detection of infection with Plasmodium parasites, and identification of insecticide resistance mechanisms. In this paper we focus on the Anopheles-specific assays that comprise the VPMT and include details of a new assay for resistance todieldrin Rdl detection. The application of these tools, general and specific guidelines on their use based on field testing in Africa, and plans for further development are discussed.

Figure 1

TaqMan SNP genotyping. The TaqMan assay is a PCR method employing oligonucleotide probes that are dual labelled with a fluorescent reporter dye and a quencher molecule. Amplification of the probe-specific product causes cleavage of the probe, generating an increase in reporter fluorescence as the reporter dye is released away from the quencher.

Figure 2

Real-time TaqMan detection of the L1014F kdr mutation. S: wild-type allele (L1014), R: resistant allele (L1014F).

Figure 3

Real-time TaqMan detection of the Rdl mutation in Anopheles gambiae and Anopheles arabiensis. (a) and (b) Detection of the A296S mutation. (c) and (d) Detection of the A296G mutation.