塔里木大学学报

研究目的:建立检测布鲁氏菌病病原巢式PCR快速诊断方法,并采用该方法进行样品检测。方法:对布鲁氏菌omp22基因设计巢式PCR引物,以布鲁氏菌基因组为模板,来检测布鲁氏菌病病原。对金黄色葡萄球菌、链球菌、沙门氏菌、大肠杆菌、棒状杆菌、绿脓杆菌、李氏杆菌等常见菌群及羊基因组进行阴性对照检测,并用本方法进行血样的检测,采用RBPT检测对应的血清样本。结果及结论:结果表明对动物的常见发病菌检测为阴性,所建立布鲁氏菌病病原PCR快速诊断方法有很好的特异性和敏感性,应用本方法对378份血样进行PCR检测,较之对应血清的RBPT检测结果,具有灵敏、特异、准确的特点,有较好的实用价值。

This study developed the nested-PCR for rapid diagnosis of brucella.Clinical samples were detected to check the practice of this method.Method: the nested-PCR primers for detecting the omp22 of brucella were designed.The genome extraction protocols of milk and blood were established and used to samples diagnosis by PCR.The Staphycococcus aureus,streptococcus,E.coli,Salmonella,Corynebacterium,Pseudomonas aeruginosa,Listeria monocytogenes were detected to take as negative control were detected by the established rapid diagnosis method.Conclusion: It is proved that the detection technique possesses stability and reproducibility by the repeated operation.The detection technique appears negative on the Staphycococcus aureus,streptococcus,E.coli,Salmonella,Corynebacterium,Pseudomonas aeruginosa,Listeria monocytogenes,which is testified that the detection technique possesses specificity and Sensitivity.We gained the ideal result in the detection of 378 samples by the nested-PCR detection method.In short,the nested-PCR for rapid diagnosis of brucella is a useful diagnosis method.