Stakenborg T, Vicca J, Butaye P, Imberechts H, Peeters J, de Kruif A, Haesebrouck F, Maes D
Veterinary and Agrochemical Research Centre, Groeselenberg 99, 1180, Brussels.
Vet Res Commun. 2006 Apr;30(3):239-47. doi: 10.1007/s11259-006-3226-3.
Mycoplasma hyopneumoniae, Mycoplasma hyorhinis and Mycoplasma flocculare can be present in the lungs of pigs at the same time. These three mycoplasma species all require similar growth conditions and can be recovered from clinical samples using the same media. We have developed a multiplex PCR as a helpful tool for rapid differentiation of these three species in the course of isolation. Based on the 16S ribosomal DNA sequences, three different forward primers and a single reverse primer were selected. Each forward primer was compared to available mycoplasma sequences, showing the primers to be specific. The three amplification products observed of 1129 bp (M. hyorhinis), 1000 bp (M. hyopneumoniae) and 754 bp (M. flocculare) were clearly distinguishable on a 1% agarose gel. In addition, no cross-reaction with Mycoplasma hyosynoviae, another porcine mycoplasma, was noted. This multiplex PCR using the proposed set of primers is the first reported assay that allows the simultaneous identification of the different Mycoplasma species isolated from the lungs of pigs.
猪肺炎支原体、猪鼻支原体和絮状支原体可同时存在于猪的肺部。这三种支原体都需要相似的生长条件，并且可以使用相同的培养基从临床样本中分离出来。我们开发了一种多重PCR技术，作为在分离过程中快速区分这三种支原体的有用工具。基于16S核糖体DNA序列，选择了三种不同的正向引物和一种反向引物。将每种正向引物与现有的支原体序列进行比较，结果表明这些引物具有特异性。在1%的琼脂糖凝胶上可以清楚地分辨出观察到的三种扩增产物，分别为1129bp（猪鼻支原体）、1000bp（猪肺炎支原体）和754bp（絮状支原体）。此外，未发现与另一种猪支原体——猪滑膜支原体有交叉反应。这种使用所提出引物组的多重PCR技术是首次报道的能够同时鉴定从猪肺部分离出的不同支原体种类的检测方法。
