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More than fishing in the dark: PCR of a dispersed sequence produces simple but ultrasensitive Wolbachia detection
Uppsala University, Disciplinary Domain of Science and Technology, Biology, Department of Cell and Molecular Biology, Molecular Evolution.
Uppsala University, Disciplinary Domain of Science and Technology, Biology, Department of Cell and Molecular Biology, Molecular Evolution.
2014 (English)In: BMC Microbiology, ISSN 1471-2180, E-ISSN 1471-2180, Vol. 14, 121- p.Article in journal (Refereed) Published
Abstract [en]

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.
Keyword [en]
Wolbachia, Drosophila, Glossina, Hybrid, High- and low-titer endosymbiont infection, Limit of detection, A-supergroup repeat motif (ARM)
National Category
Microbiology
Identifiers
URN: urn:nbn:se:uu:diva-227254DOI: 10.1186/1471-2180-14-121ISI: 000336179000001OAI: oai:DiVA.org:uu-227254DiVA: diva2:731745
Available from: 2014-07-02 Created: 2014-06-24 Last updated: 2017-12-05Bibliographically approved

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Klasson, LisaLind, Anders E.

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