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Sanidad Militar
versão impressa ISSN 1887-8571
Resumo
ORTEGA GARCIA, M.ªV. et al. Quantitative PCR in real time to Burkholderia mallei ADN amplification: A comparison with the molecular method recommended by OIE. Sanid. Mil. [online]. 2017, vol.73, n.2, pp.85-90. ISSN 1887-8571. https://dx.doi.org/10.4321/s1887-85712017000200002.
Objective:
Comparison of two quantitative real-time PCRs for identification of Burkholderia mallei, on analytical sensitivity and specificity terms.
Methods:
Partial amplification of Burkholderia mallei gene: - orf11 and orf13 targeting the type III secretion TTS1 system cluster from Burkholderia genus by qPCR using hybridisation probes. - fliP targeting flagelin P from B. mallei by qPCR using TaqMan probe.Validity parameters determination.
Results:
The duplex assay developed by the Molecular Biology Laboratory at INTA obtained a limit of detection similar than that reached by the molecular method recommended by the OIE and permitted the specific amplification of B. mallei DNA.
Conclusions:
The duplex assay developed by the Molecular Biology Laboratory at INTA provides a rapid amplification of B. mallei DNA, also shows high analytical sensitivity and specificity. Furthermore, this assay permits the differentiation between B. mallei and B. pseudomallei.
Palavras-chave : Glanders; Burkholderia mallei; Quantitative real-time PCR; Sensitivity; Specificity.