Detection and Assessment of Wolbachia pipientis Infection.

Wolbachia pipientis is a widespread vertically transmitted intracellular bacterium naturally present in the model organism Drosophila melanogaster. As Wolbachia is present in a large number of Drosophila lines, it is critical for researchers to be able to identify which of their stocks maintain this infection to avoid any potential confounding variables. Here, we describe methods for detecting the bacterium and assessing the infection, including polymerase chain reaction (PCR) of DNA, multi-locus sequence typing (MLST) to identify strains, western blotting for protein detection, and immunohistochemistry and fluorescence in situ hybridization (FISH) of Drosophila ovaries to visually detect Wolbachia by fluorescence microscopy.