Abstract
With Wolbachia-based arbovirus control programs being scaled and operationalised around the world, cost effective and reliable detection of Wolbachia in field samples and laboratory stocks is essential for quality control. Here we validate a modified loop-mediated isothermal amplification (LAMP) assay for routine scoring of Wolbachia in mosquitoes from laboratory cultures and the field, applicable to any setting. We show that this assay is a rapid and robust method for highly sensitive and specific detection of wAlbB Wolbachia infection within Aedes aegypti under a variety of conditions. We test the quantitative nature of the assay by evaluating pooled mixtures of Wolbachia-infected and uninfected mosquitoes and show that it is capable of estimating infection frequencies, potentially circumventing the need to perform large-scale individual analysis for wAlbB infection status in the course of field monitoring. These results indicate that LAMP assays are useful for routine screening particularly under field conditions away from laboratory facilities.
Highlights
Wolbachia releases are being undertaken in Aedes aegypti populations both for replacing existing populations with mosquitoes that have a reduced ability to transmit dengue [1, 2] and other arboviruses, and for suppressing mosquito populations directly due to sterility generated by males infected with Wolbachia [3]
A challenge in implementing Wolbachia-based strategies is that a high level of quality control is required for release success
It is important to track infection status in release areas given that not all releases are successful and periodic interventions may be needed for others [4, 5]
Summary
Wolbachia releases are being undertaken in Aedes aegypti populations both for replacing existing populations with mosquitoes that have a reduced ability to transmit dengue [1, 2] and other arboviruses, and for suppressing mosquito populations directly due to sterility generated by males infected with Wolbachia [3]. A challenge in implementing Wolbachia-based strategies is that a high level of quality control is required for release success. This includes ensuring that source mosquito cultures used for releases remain infected by Wolbachia. Successful replacement is dependent on Wolbachia frequencies in populations exceeding an unstable equilibrium point [1] and potential spread from a release site [6].
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