动物造模 z-bio 2022-05-06 478

　　Objective To establish a real-time, rapid, sensitive and simple PCR method for the detection of Pasteurella pneumophila in living experimental animals.

　　Methods By comparing different sample collection and processing methods, the sample processing flow was optimized, the DNA extraction steps were simplified, the PCR template was quickly obtained, and the Pasteurella pneumophila Jawetz and Heyl types were amplified by PCR, and identified by sequencing.

　　Results Among the four sampling methods, oral samples had the best detection effect; among the two DNA extraction methods, the method of boiling for 1 min after incubation could quickly obtain bacterial genomic DNA in the samples, which was better than the kit extraction method.

　　Conclusion The rapid extraction of bacterial genomic DNA from the oral samples of experimental animals by culturing and boiling for 1 min as a PCR template can significantly improve the positive detection rate of Pasteurella pneumophila.