关键词: 嗜肺巴斯德杆菌, 样本处理, DNA快速提取, PCR

Abstract:  Objective    To establish a real-time, rapid, sensitive and simple PCR method for the detection of Pasteurella pneumophila in living laboratory animals. Methods    By comparing different sample collection and processing methods, optimizing the sample processing process, and simplifying the method of DNA extraction, PCR amplification of Jawetz type and Heyl type of Pasteurella pneumophila was performed after rapid PCR template acquisition and combined with DNA sequencing for identification. Results    Among the 4 sampling methods, the oral sample had the best detection effect. In 2 methods of DNA extraction, the bacterial genomic DNA could be obtained rapidly by culturing and boiling the samples for 1 min, which was more effective than the kit extraction method. Conclusion    Rapid extraction of bacterial genomic DNA from the oral samples of experimental animals by incubation and boiling for 1 min as PCR template can significantly improve the positive detection rate of Pasteurella pneumophila.

Key words: Pasteurella pneumophila, Sample processing, Rapid DNA extraction, PCR