动物造模,PCR检测 z-bio 2021-10-20 382

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

　　Method By comparing different sample collection and processing methods, the sample processing process is optimized, the DNA extraction steps are simplified, the PCR template is quickly obtained, and the Jawetz type and Heyl type of Pasteurella pneumophila are amplified by PCR and combined with sequencing for identification.

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

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