Detection of Brucella DNA in blood samples of Marmota himalayana in Qinghai-Tibet Plateau by real-time fluorescence quantitative PCR

  Objective   To detect Brucella DNA in blood samples of Marmota himalayana in Qinghai-Tibet Plateau by real-time fluorescence quantitative PCR (real-time PCR).

  Methods   The whole blood samples of Marmota himalayana in Qinghai-Tibet Plateau were collected for serological tests of Brucella, including tiger red plate agglutination test (RBT), tube agglutination test (SAT) and colloidal gold immune test (GICA). The positive samples of serological tests were detected by real-time PCR. The SAT results was chosen as a diagnostic criteria to analyze the sensitivity and specificity of the improved degree of tube real-time PCR and evaluate the accuracy of the micro real-time PCR results.

  Results   We collected 1 466 whole blood samples of Marmota himalayana, in which 64 were positive in RBT, 28 were positive in GICA, and 18 were positive in SAT. Sixty four RBT positive samples were detected by real-time PCR, in which 56 samples and positive control strains had specific amplification fluorescence curve, 8 samples and negative control strains had no specific amplification curve. Real time PCR had a sensitivity of 100% compared with SAT and GICA and a specificity of 99.93% compared with RBT, the Kappa value was 0.81, which were highly consistent.

  Conclusion   Compared with traditional methods, real-time PCR is rapid and highly specific, which is suitable for the detection of Marmota himalayana samples.