Abstract
Effective mosquito monitoring relies on the accurate identification and characterization of the target population. Since this process requires specialist knowledge and equipment that is not widely available, automated field-deployable systems are highly desirable. We present a centrifugal microfluidic cartridge, the VectorDisk, which integrates TaqMan PCR assays in two feasibility studies, aiming to assess multiplexing capability, specificity, and reproducibility in detecting disk-integrated vector-related assays. In the first study, pools of 10 mosquitoes were used as samples. We tested 18 disks with 27 DNA and RNA assays each, using a combination of multiple microfluidic chambers and detection wavelengths (geometric and color multiplexing) to identify mosquito and malaria parasite species as well as insecticide resistance mechanisms. In the second study, purified nucleic acids served as samples to test arboviral and malaria infective mosquito assays. Nine disks were tested with 14 assays each. No false positive results were detected on any of the disks. The coefficient of variation in reproducibility tests was <10%. The modular nature of the platform, the easy adaptation of the primer/probe panels, the cold chain independence, the rapid (2–3 h) analysis, and the assay multiplexing capacity are key features, rendering the VectorDisk a potential candidate for automated vector analysis.
Highlights
IntroductionVectors are arthropod species (e.g., mosquitoes, ticks, and sandflies) that have the capability of acquiring and transmitting at least one type of vector-borne diseases, for example malaria (parasitic disease), dengue, chikungunya, West Nile, Zika, and yellow fever (viral diseases)
Vector-borne diseases make up more than 17% of all infectious diseases globally, and are responsible for more than 700,000 deaths annually, according to the World Health Organization (WHO) [1].Vectors are arthropod species that have the capability of acquiring and transmitting at least one type of vector-borne diseases, for example malaria, dengue, chikungunya, West Nile, Zika, and yellow fever
Indicative examples are: (i) the spread of the invasive Asian tiger mosquito (Aedes albopictus), which has been associated with disease transmission in north-western USA [5] and in Europe; and (ii) the continuous circulation of West Nile virus, vectored by the common mosquito Culex pipiens, which has caused epidemics with fatalities in several European countries [10,11,12]
Summary
Vectors are arthropod species (e.g., mosquitoes, ticks, and sandflies) that have the capability of acquiring and transmitting at least one type of vector-borne diseases, for example malaria (parasitic disease), dengue, chikungunya, West Nile, Zika, and yellow fever (viral diseases). Indicative examples are: (i) the spread of the invasive Asian tiger mosquito (Aedes albopictus), which has been associated with disease transmission in north-western USA [5] and in Europe (in the latter, it has been associated with several local dengue [6], chikungunya [7,8], and Zika virus [9] outbreaks); and (ii) the continuous circulation of West Nile virus, vectored by the common mosquito Culex pipiens (almost unnoticed until recently), which has caused epidemics with fatalities in several European countries [10,11,12]
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