摘要
为建立一种灵敏、特异的猪源多杀性巴氏杆菌PCR检测方法,根据GenBank已公布的多杀性巴氏杆菌plpE基因序列的保守片段设计合成引物,经plpE基因阳性质粒构建、反应条件的优化、特异性试验和敏感性试验,对猪源多杀性巴氏杆菌特异性PCR检测方法进行了研究;并将该PCR方法用于检测来自四川省6个规模化猪场的64份疑似多杀性巴氏杆菌肺部组织样品。结果显示,建立的PCR方法具有良好的特异性和敏感性,检测的敏感性为5×101拷贝的目的基因;多杀性巴氏杆菌(A、B、D血清型)PCR均为阳性、APP和HPS等病原均为阴性;64份临床疑似样品中检出36份样品阳性,阳性检出率为56.2%。以plpE基因初步建立的多杀性巴氏杆菌PCR检测方法具有较好的特异性、重复性、敏感性和可靠性,可用于多杀性巴氏杆菌的检测鉴定。
The objective of this study was to develop a sensitive and specific PCR method for the detection of Pasteurella multocida from porcine.A pair of primers were designed according to the plpE genes of Pasteurella multocida published in GenBank.Then the specific PCR method on the basis of the plpE gene was developed and optimized for rapid detection of Pasteurella multocida.The sensitivity of the method were evaluated with the constructed recombinant plasmid of Pasteurella multocida and other strains.Finally the PCR method established was employed on the detection of 64 suspected samples collected from Sichuan province.Results showed the method based on the plpE gene can amplify the genome of Pasteurella multocida only,not for others and the detection limitation reached 5×101 copies.Thirty-six samples were infected with Pasteurella multocida and the positive rate reached 56.2%.These results indicate that the PCR method on the basis of the plpE gene is specific,repeatable and sensitive and can be used for the detection of Pasteurella multocida.
出处
《畜牧兽医学报》
CAS
CSCD
北大核心
2012年第7期1111-1116,共6页
ACTA VETERINARIA ET ZOOTECHNICA SINICA
基金
教育部<长江学者和创新团队发展计划>创新团队项目(IRTO848)
四川农业大学双支计划
