论文部分内容阅读
本试验以布鲁氏菌致病力因子VirB7下游至VirB9上游基因序列为目的扩增片段,设计上、下游引物,优化血样、奶样中布鲁氏菌基因组DNA的提取方法,建立布鲁氏菌内参PCR(IR-PCR)检测方法,用于血液、奶液样本中布鲁氏菌的检测。结果显示,内参PCR法有效地降低了PCR法诊断布鲁氏菌的假阴性率,且检测结果与常规的布鲁氏菌的诊断方法(细菌培养分离鉴定、iELASA)一致,该方法不仅提高了常规布鲁氏菌诊断方法的效率及灵敏度,而且降低了其假阴性和假阳性率。对血样、奶样的检出量分别为35CFU/mL、350 CFU/mL,适合于血样、奶样中布鲁氏菌的检测。
In this study, we aimed to amplify the sequences from the virulence factor of virulent strain VirB7 to the virB9 upstream gene sequence, design the upstream and downstream primers, optimize the method of extracting the genomic DNA of Brucella from the blood sample and the milk sample, Bacterial reference PCR (IR-PCR) detection method for blood, milk samples Brucella detection. The results showed that the internal reference PCR method effectively reduced the false negative rate of PCR diagnosis of Brucella, and the test results and conventional diagnosis of Brucella (bacterial culture identification, iELASA), the method not only improves Routine Brucella diagnostic methods efficiency and sensitivity, but also reduce its false negative and false positive rate. The blood samples, the detection of milk samples were 35CFU / mL, 350 CFU / mL, suitable for blood, milk samples of Brucella detection.