Comparative application of droplet-based digital and quantitative real-time PCR for human brucellosis detection

Abstract

We evaluated the diagnostic value of droplet-based digital PCR (dd-PCR) by comparing it with the quantitative real-time PCR (RT-qPCR) for detecting Brucella DNA, 487 whole blood and serum samples collected from suspected human brucellosis, respectively. Sensitivity and specificity were 88.14% and 100% for RT-qPCR; 97.12% and 100% for dd-PCR. The positive rate detected by RT-qPCR and dd-PCR based on the nucleic acid extracted by simultaneous extraction method in serum and blood cells were 56.49% and 62.22%, respectively, which is higher than the commercial kit in 47.74% and 52.77%. Additionally, 32 false-negative samples of chronic patients analyzed by serological tests were positive in the detection from the blood cell nucleic acid. dd-PCR could be considered a valuable tool for detecting Brucella DNA, particularly in false-negative test results. The simultaneous extraction method is complementary to dd-PCR in diagnosing human brucellosis cases at different disease stages, especially in chronic and relapsed stages.