Detection, Quantification, and Elimination of Wolbachia in Bed Bugs

Wolbachia is an obligatory nutritional symbiont of the common bed bug, Cimex lectularius, providing B-vitamins to its host. The biological significance of Wolbachia to bed bugs is investigated primarily by eliminating the symbiont with antibiotics, which is followed by confirmation with molecular assays. In this chapter, we describe a protocol for eliminating Wolbachia in bed bugs using the ansamycin antibiotic rifampicin (also known as rifampin) and three molecular methods to accurately detect and quantify the Wolbachia gene copies in bed bug samples. We describe the digital droplet PCR (ddPCR), a highly sensitive technique for absolute quantification of low abundance target genes, which has proven to be a valuable technique for confirmation of the elimination of Wolbachia.