摘要
利用PCR反应扩增出胸膜肺炎放线杆菌ApxⅣ基因650 bp的DNA,回收并纯化PCR产物,用地高辛标记,制备出地高辛标记的核酸探针。该探针与不同血清型的胸膜肺炎放线杆菌均能发生特异性杂交,而与大肠杆菌、多杀性巴氏杆菌、沙门氏菌、葡萄球菌、猪肺炎支原体、支气管败血波氏杆菌等细菌的核酸杂交均为阴性。对胸膜肺炎放线杆菌的最低检出限量为10×102 个放线杆菌。对疑似胸膜肺炎放线杆菌感染病变组织检测结果表明,在扁桃体、鼻腔、气管、肺脏均可检测出胸膜肺炎放线杆菌,以扁桃体的检出率最高。为猪胸膜肺炎放线杆菌的诊断和流行病学调查提供了良好的方法。
The APXⅣ gene, a recently discovered RTX determinant of Actinobacillus pleuropneumoniae, was shown to be species-specific. A pair of primers were designed for amplifying this 650 bp fragment in PCR experiments.The PCR products was labeled with digoxigenin as DNA probe for detection of Actinobacillus pleuropneumoniae.The hybridization assay results showed that the reference strains of the 12 serotypes of Actinobacillus pleuropneumoniae were positive, no cross-hybridization was detected with DNAs from other bacterial species closely related to A. pleuropneumoniae. The sensitivity result showed that as few as 10×10 2 of Actinobacillus pleuropneumoniae could be detected by DIG-labeled probe. The detection results for clinical sample tissues showed that Actinobacillus pleuropneumoniae could be found in tonsils ,trachea and lung .The high sensitivity and specificity of the DIG-labeled probe will make it useful in field diagnostic work and epidemic investigation.
出处
《畜牧兽医学报》
CAS
CSCD
北大核心
2005年第6期585-589,共5页
ACTA VETERINARIA ET ZOOTECHNICA SINICA
基金
山东省重大畜禽疫病综合防治技术(011020102)
