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Detection and Assessment of Wolbachia pipientis Infection
Lindsay B M Nevalainen1, Irene L G Newton2
1Department of Biology, Indiana University, Bloomington, IN, USA.
Abstract:
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.
Insights
Researchers can detect Wolbachia pipientis, an intracellular bacterium in Drosophila melanogaster, using various molecular and imaging techniques. Identifying this common infection is crucial for accurate experimental results in model organism research.
Area of Science:
- Microbiology and Genetics
- Model Organism Research
Background:
- Wolbachia pipientis is a common intracellular bacterium found in Drosophila melanogaster.
- Its presence in numerous Drosophila stocks can introduce confounding variables in research.
- Accurate detection methods are essential for researchers using Drosophila melanogaster.
Purpose of the Study:
- To describe reliable methods for detecting Wolbachia pipientis infection in Drosophila melanogaster.
- To provide techniques for assessing the extent and strain of Wolbachia infection.
- To aid researchers in controlling for potential confounding factors in their experiments.
Main Methods:
- DNA analysis using polymerase chain reaction (PCR).
- Multi-locus sequence typing (MLST) for strain identification.
- Protein detection via western blotting.
- Visual detection in ovaries using immunohistochemistry and fluorescence in situ hybridization (FISH) with fluorescence microscopy.
Main Results:
- Established multiple methods for detecting Wolbachia pipientis.
- Provided techniques for strain typing and visual confirmation of infection.
- Enabled researchers to identify infected Drosophila stocks.
Conclusions:
- Multiple validated methods exist for detecting Wolbachia pipientis in Drosophila melanogaster.
- These techniques are vital for ensuring the integrity of research using this model organism.
- Researchers can confidently identify and manage Wolbachia infections in their fly stocks.
