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首页> 外文期刊>Journal of Clinical Microbiology >Evaluation of a Multiplex PCR Test for Simultaneous Identification and Serotyping of Actinobacillus pleuropneumoniae Serotypes 2, 5, and 6
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Evaluation of a Multiplex PCR Test for Simultaneous Identification and Serotyping of Actinobacillus pleuropneumoniae Serotypes 2, 5, and 6

机译：同时鉴定和鉴定血清胸膜肺炎放线杆菌血清型2、5和6的多重PCR试验的评价

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Serotype-specific DNA regions involved in the biosynthesis of capsular polysaccharides (cps region) were used to develop a multiplex PCR test for the simultaneous species identification and serotyping of Actinobacillus pleuropneumoniae serotypes 2, 5, and 6. Primers specific for serotypes 2, 5, and 6 were combined with the already existing species-specific primers used in a PCR test based on the omlA gene. The PCR test was evaluated with serotype reference strains of A. pleuropneumoniae as well as 182 Danish field isolates previously serotyped by latex agglutination or immunodiffusion. For all serologically typeable strains, a complete correspondence was found between the results obtained by the multiplex PCR test and the results obtained by the traditional serotyping methods. Six of eight serologically nontypeable strains could be allocated to a serotype on the basis of the multiplex PCR results. The species specificity of the assay was evaluated with a collection of 93 strains representing 29 different species within the family Pasteurellaceae, as well as species normally found in the respiratory tracts of swine. All of these strains were negative by the multiplex PCR test, including 50 field isolates of the phylogenetically closely related species Actinobacillus lignieresii. When the multiplex PCR test was used to test Danish field strains, it was able to identify the serotypes of approximately 94% of all strains isolated from swine with clinical disease. More than 90% of the isolates that cross-reacted by the latex agglutination test were of serotype 2, 5, or 6. Determination of the serotype by PCR represents a convenient and specific method for the serotyping of A. pleuropneumoniae in diagnostic laboratories.
机译：利用参与荚膜多糖生物合成的血清型特异性DNA区域（ cps 区域）开发了多重PCR试验，用于同时鉴别胸膜肺炎放线杆菌（eminobacillus pleuropneumoniae）血清型2的种类和血清分型。 ，5和6。针对血清型2、5和6的引物与基于 omlA 基因进行PCR检测的物种特异性引物结合在一起。用血清型参考菌株A评估PCR试验。胸膜肺炎以及以前通过乳胶凝集或免疫扩散进行血清分型的182种丹麦田间分离株。对于所有血清学可分型的菌株，在多重PCR测试获得的结果与传统血清分型方法获得的结果之间找到了完全对应的关系。根据多重PCR结果，可以将八种血清学不可分型的菌株中的六株分配为血清型。共有93个品系代表了该测定的物种特异性，这些品系代表巴氏杆菌科中的29个不同物种，以及通常在猪呼吸道中发现的物种。通过多重PCR测试，所有这些菌株均为阴性，包括系统发育上密切相关的物种 Actinobacillus lignieresii 的50个现场分离株。当使用多重PCR测试来测试丹麦田间菌株时，它能够鉴定出从患有临床疾病的猪中分离出的所有菌株中约94％的血清型。通过乳胶凝集试验交叉反应的分离株中，有90％以上是血清型2、5或6。通过PCR确定血清型代表了一种简便，特异的 A血清分型方法。胸膜肺炎在诊断实验室。

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