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Viruses 2017, 9(6), 144; doi:10.3390/v9060144

Highly Sensitive Bacteriophage-Based Detection of Brucella abortus in Mixed Culture and Spiked Blood

Department of Bacteriophage Therapeutics, Bacterial Diseases Branch, Walter Reed Army Institute of Research, Silver Spring, MD 20910, USA
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Academic Editors: Tessa E. F. Quax, Matthias G. Fischer and Laurent Debarbieux
Received: 1 April 2017 / Revised: 25 May 2017 / Accepted: 6 June 2017 / Published: 10 June 2017
(This article belongs to the Special Issue Viruses of Microbes)
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Abstract

For decades, bacteriophages (phages) have been used for Brucella species identification in the diagnosis and epidemiology of brucellosis. Traditional Brucella phage typing is a multi-day procedure including the isolation of a pure culture, a step that can take up to three weeks. In this study, we focused on the use of brucellaphages for sensitive detection of the pathogen in clinical and other complex samples, and developed an indirect method of Brucella detection using real-time quantitative PCR monitoring of brucellaphage DNA amplification via replication on live Brucella cells. This assay allowed the detection of single bacteria (down to 1 colony-forming unit per milliliter) within 72 h without DNA extraction and purification steps. The technique was equally efficient with Brucella abortus pure culture and with mixed cultures of B. abortus and α-proteobacterial near neighbors that can be misidentified as Brucella spp., Ochrobactrum anthropi and Afipia felis. The addition of a simple short sample preparation step enabled the indirect phage-based detection of B. abortus in spiked blood, with the same high sensitivity. This indirect phage-based detection assay enables the rapid and sensitive detection of live B. abortus in mixed cultures and in blood samples, and can potentially be applied for detection in other clinical samples and other complex sample types. View Full-Text
Keywords: Brucella abortus; brucellosis; phage-based detection; phage amplification; qPCR; rapid assay; blood and clinical samples Brucella abortus; brucellosis; phage-based detection; phage amplification; qPCR; rapid assay; blood and clinical samples
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MDPI and ACS Style

Sergueev, K.V.; Filippov, A.A.; Nikolich, M.P. Highly Sensitive Bacteriophage-Based Detection of Brucella abortus in Mixed Culture and Spiked Blood. Viruses 2017, 9, 144.

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