Optimisation of detection methods for Wolbachia bacterial strains in Anopheles mosquitoes

Wolbachia bacteria can invade mosquito populations and inhibit pathogen transmission. Wolbachia strains have recently been shown to be in genuine endosymbiosis in two Anopheles mosquito species (the vectors of malaria). Current PCR-based detection likely underestimates the prevalence and diversity of strains in wild Anopheles populations. The aim of this Phd is to develop, optimise and validated PCR and sequencing based Wolbachia detection methods suitable for low-resource settings. The PhD will utilise existing biobanks of Anopheles mosquitoes collected from malaria-endemic WHO Africa region countries such as Kenya and the Democratic Republic of Congo to firstly determine the optimal methods for both mosquito DNA extraction and PCR-based detection of Wolbachia genes. The optimised methodology will be tested using newly collected mosquito samples from fieldwork undertaken in Kissi, Kenya. The project will then determine the limits of detection for Wolbachia strains and determine if downstream illumina and oxford nanopore technologies (ONT) sequencing can be used to characterise both existing and newly discovered Wolbachia strains in Anopheles mosquitoes.