Development of a multiplex PCR test for identification of Actinobacillus pleuropneumoniae serovars 1, 7, and 12
Development of a multiplex PCR test for identification of Actinobacillus pleuropneumoniae serovars 1, 7, and 12
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DOI：
10.1016/j.vetmic.2008.05.010
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发表时间：
2008-12-10
影响因子：
3.3
通讯作者：
Jessing, Stine G.
中科院分区：
文献类型：
--
作者：
Angen, Oystein;Ahrens, Peter;Jessing, Stine G.
开发了一种用于同时鉴定和分离胸膜肺炎放线杆菌血清型 1、7 和 12 的 PCR 测定方法。将参与血清型 1、7 和 12 荚膜多糖（cps 基因）生物合成的基因特异的引物与基于 omlA 基因的物种特异性 PCR 测试相结合。 PCR 测试使用胸膜肺炎放线菌血清型参考菌株以及 183 个丹麦野外分离株进行评估。对于所有可分型的菌株，多重 PCR 测试获得的结果与传统血清分型方法的结果之间存在完全对应。在指定为 K 1:07 的 8 个血清学交叉反应菌株中，7 个分离株产生与血清型 I 大小相似的扩增子，1 个分离株产生与血清型 7 大小相似的扩增子。使用代表巴斯德氏菌科内 25 个不同物种的 126 个菌株（包括系统发育附属物种放线杆菌的 45 个野外菌株）评估了测定的物种特异性。 利涅雷西。通过多重 PCR 测试，除 6 个 A. lignieresii 分离株外，所有这些分离株的 cps 基因均呈阴性。这些分离株中的五个产生了与血清型7的cps基因相同的扩增子，而一个分离株产生了与血清型1的cps基因相同的所有扩增子。放线杆菌基因种 1 的四个分离株的 omlA 基因检测呈阳性，但 cps 基因呈阴性。该测试代表了诊断实验室中对胸膜肺炎放线菌进行血清分型的一种方便且特异的方法。 (C) 2008 Elsevier B.V. 保留所有权利。
A PCR assay for simultaneous species identification and separation of Actinobacillus pleuropneumoniae serovars 1, 7 and 12 was developed. Primers specific for genes involved in biosynthesis of the capsular polysaccharides (cps genes) of serovars 1, 7, and 12 were combined with a species-specific PCR test based on the omlA gene. The PCR test was evaluated with the serovar reference strains of A. pleuropneumoniae as well as 183 Danish field isolates. For all typable strains, a complete correspondence was found between results obtained with the multiplex PCR test and results from the traditional serotyping methods. Among eight serologically cross-reacting strains designated K 1:07, seven isolates produced amplicons of similar sizes as serovar I and one isolate produced amplicons of similar sizes as serovar 7. The species specificity of the assay was evaluated using a collection of 126 strains representing 25 different species within the family Pasteurellaceae including 45 field strains of the phylogenetically affiliated species Actinobacillus lignieresii. All these isolates tested negative for the cps genes by the multiplex PCR test except for 6 isolates of A. lignieresii. Five of these isolates produced an amplicon identical to the cps gene of serovar 7, whereas one isolate produced ail amplicon identical to the cps gene of serovar 1. In addition. four isolates of Actinobacillus genomospecies 1 tested positive for the omlA gene but negative for the cps genes. The test represents a convenient and specific method for serotyping A. pleuropneumoniae in diagnostic laboratories. (C) 2008 Elsevier B.V. All rights reserved.