实时荧光定量PCR检测布鲁氏菌核酸DNA的应用评价

目的探讨实时荧光定量PCR检测布鲁氏菌核酸DNA浓度的检测范围和检测健康人全血中核酸DNA的Ct本底值。方法采用试剂盒提取纯菌、健康人和临床患者血液中的核酸DNA，应用实时荧光定量PCR进行检测。结果经检测，40份健康人群血液标本Ct值平均值为37.0，标准差为1.9，Ct值95%置信区间为33.0 ~ 38.8。 将布鲁氏菌核酸DNA标准品倍比稀释（56.2 ng ~ 0.026 5 fg），实时荧光定量PCR检测灵敏度为6.7 fg。结论实时荧光定量PCR检测纯菌核酸DNA的灵敏度相当于2个基因组DNA拷贝数，但用于检测血液标本尚待进一步研究。

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.