Abstract
BackgroundThe World Mosquito Program uses Wolbachia pipientis for the biocontrol of arboviruses transmitted by Aedes aegypti mosquitoes. Diagnostic testing for Wolbachia in laboratory colonies and in field-caught mosquito populations has typically employed PCR. New, simpler methods to diagnose Wolbachia infection in mosquitoes are required for large-scale operational use.MethodsField-collected Ae. aegypti mosquitoes from North Queensland were tested using primers designed to detect the Wolbachia wsp gene, specific to the strain wMel. The results were analysed by colour change in the reaction mix. Furthermore, to confirm the efficiency of the LAMP assay, the results were compared to the gold-standard qPCR test.ResultsA novel loop-mediated isothermal amplification (LAMP) colorimetric test for the wMel strain of Wolbachia was designed, developed and validated for use in a high-throughput setting. Against the standard qPCR test, the analytical sensitivity, specificity and diagnostic metrics were: sensitivity (99.6%), specificity (92.2%), positive predictive value (97.08%) and negative predictive value (99.30%).ConclusionsWe describe an alternative, novel and high-throughput method for diagnosing wMel Wolbachia infections in mosquitoes. This assay should support Wolbachia surveillance in both laboratory and field populations of Ae. aegypti.
Highlights
The World Mosquito Program uses Wolbachia pipientis for the biocontrol of arboviruses transmitted by Aedes aegypti mosquitoes
Numerous studies demonstrated that the presence of wMel reduces dengue virus [9], Zika [10], chikungunya virus [11] yellow fever [11] and Mayaro virus [12] infection and replication and, in turn the virus transmission potential of the mosquito [13, 14]. wMel has been established in field populations of Ae. aegypti in five countries and it Gonçalves et al Parasites Vectors (2019) 12:404 has been expanded to more than ten worldwide [15]
Equivocal loop-mediated isothermal amplification (LAMP) results were more likely to result from qPCR-negative samples than qPCR-positive samples
Summary
The World Mosquito Program uses Wolbachia pipientis for the biocontrol of arboviruses transmitted by Aedes aegypti mosquitoes. Arboviral diseases transmitted by the mosquito Aedes aegypti such as dengue, chikungunya and Zika constitute a significant burden to human health and economic development worldwide [1, 2]. This is reflected in the nomination by the World Health Organisation of dengue as one of the top ten global health threats in 2019. Worldmosquitoprogram.org, formerly known as the Eliminate Dengue Program) has developed a novel arboviral disease biocontrol strategy utilising the endosymbiotic bacterium Wolbachia pipientis. This maternally transmitted bacterium [4] is found in 40–60% of insect species worldwide [5,6,7]. The mitigation of local dengue outbreaks in northern Australia [16] following the establishment of wMel is consistent with the laboratory and modelling expectations of this intervention [17]
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