Development of a specific real-time PCR assay for simultaneous detection and differentiation of Coxiella burnetii strains from environmental soil samples.

Autor:	Fernández-Carrillo J; Biological Defence Area, CBRN Defence Systems Department, Campus La Marañosa, Instituto Nacional de Técnica Aeroespacial (INTA), San Martín de la Vega, 28330 Madrid, Spain., Del Olmo-Monge J; Biological Defence Area, CBRN Defence Systems Department, Campus La Marañosa, Instituto Nacional de Técnica Aeroespacial (INTA), San Martín de la Vega, 28330 Madrid, Spain., Sellek RE; Biological Defence Area, CBRN Defence Systems Department, Campus La Marañosa, Instituto Nacional de Técnica Aeroespacial (INTA), San Martín de la Vega, 28330 Madrid, Spain., Ortega-García MV; Biological Defence Area, CBRN Defence Systems Department, Campus La Marañosa, Instituto Nacional de Técnica Aeroespacial (INTA), San Martín de la Vega, 28330 Madrid, Spain., Cabria-Ramos JC; Biological Defence Area, CBRN Defence Systems Department, Campus La Marañosa, Instituto Nacional de Técnica Aeroespacial (INTA), San Martín de la Vega, 28330 Madrid, Spain., Bassy O; Biological Defence Area, CBRN Defence Systems Department, Campus La Marañosa, Instituto Nacional de Técnica Aeroespacial (INTA), San Martín de la Vega, 28330 Madrid, Spain.
Jazyk:	angličtina
Zdroj:	Letters in applied microbiology [Lett Appl Microbiol] 2023 Mar 01; Vol. 76 (3).
DOI:	10.1093/lambio/ovad030
Abstrakt:	Coxiella burnetii, the causative agent of Q fever, is a small, coccoid, Gram-negative strict intracellular pathogen. One of the most common ways of acquiring Q fever is through inhalation of aerosols containing the bacteria. Because C. burnetii is highly infectious, spreads easily through the air, and is very resistant to environmental conditions, it is considered a biological threat. This paper presents the development and validation of a specific real-time polymerase chain reaction (real-time PCR or qPCR) assay for the detection of C. burnetii, based on the amplification of a fragment of the isocitrate dehydrogenase (icd) encoding gene. This real-time PCR is highly specific, reproducible, and sensitive, allowing the detection of as few as 5 genome equivalents (GEs) of C. burnetii per reaction. The method enables a rapid preliminary differentiation among strains, based on a point mutation at nucleotide 745 of the icd gene. The assay was successfully evaluated in environmental soil samples; a limit of detection of 3 × 104 colony forming units per 0.5 g of soil (∼3 GEs per reaction) was achieved. The newly developed real-time PCR offers a valuable tool for differential detection of C. burnetii strains in environmental soil samples. (© The Author(s) 2023. Published by Oxford University Press on behalf of Applied Microbiology International.)
Databáze:	MEDLINE
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