More than fishing in the dark: PCR of a dispersed sequence produces simple but ultrasensitive Wolbachia detection
2014 (English)In: BMC Microbiology, ISSN 1471-2180, Vol. 14, 121- p.Article in journal (Refereed) Published
Background: Detecting intracellular bacterial symbionts can be challenging when they persist at very low densities. Wolbachia, a widespread bacterial endosymbiont of invertebrates, is particularly challenging. Although it persists at high titers in many species, in others its densities are far below the detection limit of classic end-point Polymerase Chain Reaction (PCR). These low-titer infections can be reliably detected by combining PCR with DNA hybridization, but less elaborate strategies based on end-point PCR alone have proven less sensitive or less general. Results: We introduce a multicopy PCR target that allows fast and reliable detection of A-supergroup Wolbachia -even at low infection titers -with standard end-point PCR. The target is a multicopy motif (designated ARM: A-supergroup repeat motif) discovered in the genome of wMel (the Wolbachia in Drosophila melanogaster). ARM is found in at least seven other Wolbachia A-supergroup strains infecting various Drosophila, the wasp Muscidifurax and the tsetse fly Glossina. We demonstrate that end-point PCR targeting ARM can reliably detect both high-and low-titer Wolbachia infections in Drosophila, Glossina and interspecific hybrids. Conclusions: Simple end-point PCR of ARM facilitates detection of low-titer Wolbachia A-supergroup infections. Detecting these infections previously required more elaborate procedures. Our ARM target seems to be a general feature of Wolbachia A-supergroup genomes, unlike other multicopy markers such as insertion sequences (IS).
Place, publisher, year, edition, pages
2014. Vol. 14, 121- p.
Wolbachia, Drosophila, Glossina, Hybrid, High- and low-titer endosymbiont infection, Limit of detection, A-supergroup repeat motif (ARM)
IdentifiersURN: urn:nbn:se:uu:diva-227254DOI: 10.1186/1471-2180-14-121ISI: 000336179000001OAI: oai:DiVA.org:uu-227254DiVA: diva2:731745