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取病死鸡肝接种于血清琼脂平板,挑取符合巴氏杆菌特征的单菌落进行纯化培养和染色镜检;对纯培养菌提取核酸,进行16S r RNA、kmt1和cap A基因的PCR鉴定,7个管家基因的PCR扩增和多位点序列分型,以及12个毒力基因的PCR检测;并进行分离菌的动物回归试验。结果显示,分离的2株病原菌(HN-1和CZ-6)均为两极浓染的革兰阴性短杆菌,16S r RNA和kmt1基因序列与多杀性巴氏杆菌的同源性均高于99%,荚膜血清型鉴定为A型,基因型为ST129型,除tox A、ptf A、nan H之外的9个毒力基因均被检出,动物回归试验表明分离菌对鸡的致病性较强。结果表明,这2株鸡源多杀性巴氏杆菌的分离鉴定为我国禽源菌株的遗传特性研究提供了一定参考。
The diseased chickens were inoculated on serum agar plates, single colony characterized by Pasteurellis was picked for purification and microscopic examination. Nucleic acids were extracted from pure cultures and identified by PCR of 16S rRNA, kmt1 and cap A genes. 7 PCR amplification of a housekeeping gene and multi-site sequence typing, and PCR detection of 12 virulence genes; and the animal regression test of the isolated bacteria. The results showed that two isolates of pathogenic bacteria (HN-1 and CZ-6) were both bipolar-stained Gram-negative Brevibacteria. The sequences of 16S rRNA and kmt1 shared more homology with Pasteurella multocida 99%, capsular serotypes identified as type A, genotype ST129, except for tox A, ptf A, nan H outside of the nine virulence genes were detected, animal regression test showed that the isolation of chickens caused by bacteria Very sick. The results showed that the isolation and identification of two strains of Pasteurella multocida were a reference for the study of the genetic characteristics of poultry origin strains in China.