Multiplex real-time PCR for detection of anaplasma phagocytophilum and Borrelia burgdorferi

Journal of Clinical Microbiology
Joshua W CourtneyRobert F Massung

Abstract

A multiplex real-time PCR assay was developed for the simultaneous detection of Anaplasma phagocytophilum and Borrelia burgdorferi. The assay was tested on various Anaplasma, Borrelia, Erhlichia, and Rickettsia species, as well as on Bartonella henselae and Escherichia coli, and the assay was found to be highly specific for A. phagocytophilum and the Borrelia species tested (B. burgdorferi, B. parkeri, B. andersonii, and B. bissettii). The analytical sensitivity of the assay is comparable to that of previously described nested PCR assays (A. phagocytophilum, 16S rRNA; B. burgdorferi, fla gene), amplifying the equivalent of one-eighth of an A. phagocytophilum-infected cell and 50 borrelia spirochetes. The dynamic range of the assay for both A. phagocytophilum and B. burgdorferi was >/=4 logs of magnitude. Purified DNA from A. phagocytophilum and B. burgdorferi was spiked into DNA extracted from uninfected ticks and from negative control mouse and human bloods, and these background DNAs were shown to have no significant effect on sensitivity or specificity of the assay. The assay was tested on field-collected Ixodes scapularis ticks and shown to have 100% concordance compared to previously described non-probe-based PCR assays. To...Continue Reading

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Citations

Jul 6, 2011·Experimental & Applied Acarology·Vincenzo LorussoDomenico Otranto
Nov 20, 2013·Ticks and Tick-borne Diseases·Valentina CapliginaRenate Ranka
Mar 15, 2011·Parasite : Journal De La Société Française De Parasitologie·C SilaghiK Pfister
Jan 18, 2013·The New England Journal of Medicine·Joseph L GugliottaSam R Telford
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Jan 6, 2012·Vector Borne and Zoonotic Diseases·Hongtao JinJun Qian
Nov 22, 2012·Vector Borne and Zoonotic Diseases·Dennis R BellSusan Sanchez
Apr 18, 2013·Vector Borne and Zoonotic Diseases·Julia GellerIrina Golovljova
Feb 21, 2013·Vector Borne and Zoonotic Diseases·Judy BettridgeRichard J Birtles
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Jul 19, 2005·Emerging Infectious Diseases·Sigurdur SkarphédinssonKåre Kristiansen

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