Wolbachia is a naturally occurring bacterium that is present in up to 60% of insect species in the environment. It is a genus of gram-negative bacteria and is safe for humans and animals. It infects most arthropods but not the Aedes aegypti, which is the primary species responsible for transmitting human viruses such as dengue, chikungunya, and Zika. Research has shown that when Wolbachia is introduced into Aedes aegypti mosquitoes, it stops these viruses from growing and being transmitted to people. This important discovery has the potential to transform the fight against life-threatening viral diseases. Wolbachia mosquitoes are now being released for virus control. To assess the performance of these measures, new diagnostic tools are required to monitor both virus and Wolbachia presence within a wild mosquito population. To enable cost-effective detection within this context, we propose the development of a novel method to assess levels of both viral and Wolbachia antigens within a mosquito sample. To facilitate this goal, we generated anti Wolbachia polyclonal sera using a recombinant Wolbachia surface protein (WSP) as an antigen. WSP, with a molecular weight of 24 kDa, was expressed using a bacterial expression system and purified under denaturing conditions via immobilized metal affinity chromatography. Purified proteins were used to immunized mice and rabbits with the recombinant proteins and generated polyclonal antibodies. We also harvested native Wolbachia proteins from JW18 cell lines to assess native WSP reactivity. Antibody reactivity was detected using the polyclonal and commercial Wolbachia antibodies against the recombinant and native Wolbachia, including homogenizing mosquitoes from the wild. These reagents will now be used to develop a novel WSP based detection method for Wolbachia detection.