巴斯德杆菌属CODEHOP PCR检测方法的建立与初步应用
发布时间：2018-09-04 11:47
【摘要】：目的建立巴斯德杆菌的CODEHOP PCR快速检测方法,为实验动物的呼吸道细菌的控制提供参考。方法应用CODEHOP在线简并引物设计工具,比对Genbank中13株巴斯德杆菌的RNA聚合酶β亚基(rpo B)氨基酸序列设计简并引物。对建立CODEHOP PCR方法用21株参考菌株进行特异性和敏感性评价,并应用于实验动物中的巴斯德杆菌检测。结果简并引物Past F6/PastR5扩增标准菌株的目的片段为200 bp左右。能够区分受试的巴斯德杆菌和主要的实验动物呼吸道病原菌。敏感性为0.2 pg/μL~2 pg/μL。在受试的609只实验动物中呼吸道样品中检测出巴斯德杆菌阳性率为19.1%。样品阳性片段经测序验证,准确率为100%。结论所建立的方法具有良好的特异性和敏感性,可用于动物样品中巴斯德杆菌的检测。
[Abstract]:Objective to establish a rapid CODEHOP PCR method for the detection of Pasteurella vulgaris and to provide reference for the control of respiratory tract bacteria in laboratory animals. Methods the degenerate primers of RNA polymerase 尾 subunit (rpo B) of 13 Pasteurella strains in Genbank were designed by using CODEHOP online degenerate primer design tool. The CODEHOP PCR method was used to evaluate the specificity and sensitivity of 21 reference strains and was applied to the detection of Pasteurella in laboratory animals. Results the target fragment of standard strain amplified by degenerate primer Past F6/PastR5 was about 200 bp. Pasteurella can be distinguished from the main laboratory animals respiratory pathogens. The sensitivity was 0.2 pg/ 渭 L ~ (2) pg/ 渭 L. The positive rate of Pasteurella was 19.1 in the respiratory tract samples of 609 experimental animals. The positive fragment was confirmed by sequencing and the accuracy was 100%. Conclusion the method has good specificity and sensitivity and can be used for the detection of Pasteurella in animal samples.
【作者单位】： 中国食品药品检定研究院实验动物资源研究所;中国医学科学院/北京协和医学院医学生物学研究所树，
本文编号：2222008
[Abstract]:Objective to establish a rapid CODEHOP PCR method for the detection of Pasteurella vulgaris and to provide reference for the control of respiratory tract bacteria in laboratory animals. Methods the degenerate primers of RNA polymerase 尾 subunit (rpo B) of 13 Pasteurella strains in Genbank were designed by using CODEHOP online degenerate primer design tool. The CODEHOP PCR method was used to evaluate the specificity and sensitivity of 21 reference strains and was applied to the detection of Pasteurella in laboratory animals. Results the target fragment of standard strain amplified by degenerate primer Past F6/PastR5 was about 200 bp. Pasteurella can be distinguished from the main laboratory animals respiratory pathogens. The sensitivity was 0.2 pg/ 渭 L ~ (2) pg/ 渭 L. The positive rate of Pasteurella was 19.1 in the respiratory tract samples of 609 experimental animals. The positive fragment was confirmed by sequencing and the accuracy was 100%. Conclusion the method has good specificity and sensitivity and can be used for the detection of Pasteurella in animal samples.
【作者单位】： 中国食品药品检定研究院实验动物资源研究所;中国医学科学院/北京协和医学院医学生物学研究所树，
本文编号：2222008
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