Application of real-time fluorescence quantitative PCR to detect DNA of Brucella

ObjectiveTo understand the detection range of Brucella nucleic acid DNA concentration and the background Ct value of nucleic acid DNA in whole blood from healthy people by using fluorescence quantitative PCR.

MethodsGenomic DNA extraction kits were used to extract nucleic acid DNA from pure Brucella, the blood samples of healthy people and clinical patients. Real-time fluorescence quantitative PCR was used to detect the nucleic acid DNA.

ResultsThe average Ct value of 40 healthy blood samples was 37.0. The standard deviation was 1.9. The 95% range of Ct value was 33.0–38.8. The sensitivity of real-time fluorescence quantitative PCR was 6.7 fg in the light of double dilution of extracted Brucella nucleic acid DNA (56.2 ng–0.026 5 fg).

ConclusionThe sensitivity of real-time fluorescence quantitative PCR to detect nucleic acid DNA from pure Brucella was equal to 2 genomic DNA copies.