二温式PCR检测猪传染性胸膜肺炎放线杆菌方法的建立与应用

为了探索简便、快速确诊猪传染性胸膜肺炎的方法,根据猪传染性胸膜肺炎放线杆菌(APP)apx IV基因序列,设计合成1对特异性引物,建立聚合酶链反应(PCR)检测APP的方法。采用该方法对APP和其他6种猪病病原核酸进行检测,结果只对APP扩增出与预期大小相符的422bp DNA片段,而对其他6种猪病病原核酸的扩增结果为阴性。该PCR最低可检出10pg的APP DNA。对送检的46份可疑病猪组织进行检测,结果有19份样品为阳性,27份为其他病原感染。

Establishment and application of a two-temperature polymerase chain reaction on detection of pig Actinobacillus pleuropneumoniae

In order to find a simple and quick method to detect porcine Actinobacillus pleuropneumoniae,a pair of primer was designed according to the sequence of apxIV gene of APP,and a two-temperature polymerase chain reaction(PCR) method was developed for the detection of porcine Actinobacillus pleuropneumoniae.Using the method to detect APP strains and nucleic acid of six porcine pathogens,the DNA fragment with a 422 bp-long was amplified from APP strains,but the other was negative with no amplified fragment.As little as 10 pg of APP DNA was detected by this PCR.Using this method to diagnose APP in clinical specimens in Guangxi,the results indicated that 19 of 46 specimens from pig with respiratory syndromes were positive and infected with APP,others were infected with the other pathogens.