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KMID : 0359320150550020105
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Korean Journal of Veterinary Research
2015 Volume.55 No. 2 p.105 ~ p.110
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Identification of Brucella melitensis isolates originating from Mongolia and diagnostic real-time PCR evaluation using a specific SNP
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Kang Sung-Il
Kim Ji-Yeon
Kim Suk-Mi
Lee Jin-Ju
Sung So-Ra
Kim Yeon-Hee
Jung Suk-Chan
Her Moon
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Abstract
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A real-time PCR assay using hybridization probe (HybProbe) has been developed to detect Brucella (B.) melitensis strains. The primer and HybProbe sets were designed based on the gap gene of chromosome I with a specific single nucleotide polymorphism of B. melitensis. Specificity of the assay was confirmed by comparison to reference Brucella species and other related strains. In the melting curve analysis, B. melitensis generated a peak at 67oC unlike those for other Brucella species observed at 61oC. Sensitivity of the assay for B. melitensis ranged from 20 ng to 200 fg of genomic DNA. The ability to identify 94 Mongolian B. melitensis isolates using the real-time PCR assay was identical to that of classical biotyping methods and differential multiplex PCR. These data showed that this new molecular technique is a simple and quick method for detecting B. melitensis, which will be important for the control and prevention of brucellosis.
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KEYWORD
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Brucella melitensis, HybProbe, melting curve, real-time PCR
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