猪胸膜肺炎放线杆菌PCR快速分型系统的建立及应用

根据外膜蛋白基因中间部分的差异,毒素基因和荚膜基因的型特异性,建立3个多重PCR的分型体系,可将App1～12型分成11个模式,除9型和11型外完全区分。此分型体系用于3株野毒株的分型,得出的结果和血清学分型一致。对人工发病猪扁桃体、肺脏、鼻拭子各12份分型结果与已知血清型一致。将该分型系统直接用于临床病料的鉴定并分型,实验从35份临床可疑病料中检测出2份阳性,均为7型,从健康猪扁桃体350份中检测出5份阳性,一份为4型,一份为12型,一份为3型,2份为7型。

Development of a PCR Rapid Typing System for Identification and Serotyping of Actinobacillus Pleuropneumoniae and Its Application

With the variable middle region of the omlA gene and the specificlty of the apx gene and the cps gene, a typing system of three multiplex PCRs was established. By the PCR system, 11 out of the 12 reference strains of Actinobacillus pleuropneumoniae were differentiated except distinguishing serotype 9 from 11. Three wild strains serotyped from the serological tests were consistent with the typing system serotyping results, and the serotyping results of artificially infected specimens were also accurate, including 12 tonsils, 12 lungs, 12 nasal swabs. The method can also be used directly to detect A. pleuropneumoniae from the clinical suspicion samples, and 2 positive results detected from 35 clinical suspicion samples were both ascribed to serotype 7.And 5 positive results detected from 350 tonsils of healthy swine were serotype 4, serotypel2, serotype3 and two serotype7.