A novel real-time PCR assay for specific detection of Brucella melitensis

BMC Infectious Diseases

Research Article

Rene Kaden¹  Erik Alm²  Sevinc Ferrari³  Tara Wahab⁴ 

[1]Department of Medical Sciences, Clinical Microbiology, Uppsala University, Uppsala, Sweden

[2]Department of Microbiology, The Public Health Agency of Sweden, Stockholm, Sweden

[3]National Veterinary Institute, Uppsala, Sweden

[4]Swedish Forum for Biopreparedness Diagnostics, Stockholm, Umeå and Uppsala, Sweden

[5]Swedish Forum for Biopreparedness Diagnostics, Stockholm, Umeå and Uppsala, Sweden

[6]Department of Microbiology, The Public Health Agency of Sweden, Stockholm, Sweden

关键词: Brucella melitensis;    Brucellosis;    Real-Time PCR;   

DOI  :  10.1186/s12879-017-2327-7

 received in 2016-11-16, accepted in 2017-03-16,  发布年份 2017

来源: Springer

【 摘 要 】

BackgroundBrucellosis is a zoonosis that occurs worldwide. The disease has been completely eradicated in livestock in Sweden in 1994, and all cases of confirmed human brucellosis are imported into Sweden from other countries. However, due to an increase in the number of refugees and asylum seekers from the middle-east to Sweden, there is a need to improve the current diagnostic methodology for Brucella melitensis. Whilst culture of Brucella species can be used as a diagnostic tool, real-time PCR approaches provide a much faster result. The aim of this study was to set up a species-specific real-time PCR for the detection of all biovars of Brucella melitensis, which could be used routinely in diagnostic laboratories.MethodsA Brucella melitensis real-time PCR assay was designed using all available genomes in the public database of Brucella (N = 96) including all complete genomes of Brucella melitensis (N = 17). The assay was validated with a collection of 37 Brucella species reference strains, 120 Brucella melitensis human clinical isolates, and 45 clinically relevant non-Brucella melitensis strains.ResultsIn this study we developed a single real-time PCR for the specific detection of all biovars of Brucella melitensis.ConclusionsThis new real-time PCR method shows a high specificity (100%) and a high sensitivity (1.25 GE/μl) and has been implemented in the laboratories of four governmental authorities across Sweden.

【 授权许可】

CC BY   
© The Author(s). 2017