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通过体外扩增16S rRNA基因检测猪鼻拭子中的猪肺炎支原体。

Detection of Mycoplasma hyopneumoniae in nose swabs from pigs by in vitro amplification of the 16S rRNA gene.

作者信息

Mattsson J G, Bergström K, Wallgren P, Johansson K E

机构信息

National Veterinary Institute, Uppsala, Sweden.

出版信息

J Clin Microbiol. 1995 Apr;33(4):893-7. doi: 10.1128/jcm.33.4.893-897.1995.

Abstract

We report the use of PCR to detect DNA from Mycoplasma hyopneumoniae, the etiological agent of enzootic porcine pneumonia. A primer set was designed for the amplification of a 649-bp fragment of the 16S rRNA gene from M hyopneumoniae. The PCR product was identified by ethidium bromide staining after gel electrophoresis and by Southern hybridization with an M. hyopneumoniae-specific oligonucleotide probe. No amplification was observed from any other porcine bacteria tested, including several closely related mycoplasmas. It was also possible to demonstrate the presence of M. hyopneumoniae in bronchial lavage samples and lung tissue samples from experimentally infected pigs. Furthermore, the PCR system was used for analysis of nasal samples obtained from pigs in a fattening herd. By this method, we were able to detect M. hyopneumoniae in nose swabs from naturally infected pigs. However, our results suggest that M. hyopneumoniae can be detected in the nasal cavities only during a limited time period.

摘要

我们报告了使用聚合酶链反应(PCR)检测猪肺炎支原体DNA的情况,猪肺炎支原体是地方性猪肺炎的病原体。设计了一组引物,用于扩增猪肺炎支原体16S rRNA基因的一个649碱基对片段。PCR产物经凝胶电泳后用溴化乙锭染色,并与猪肺炎支原体特异性寡核苷酸探针进行Southern杂交来鉴定。在所检测的任何其他猪细菌中均未观察到扩增,包括几种密切相关的支原体。还能够证明在实验感染猪的支气管灌洗样本和肺组织样本中存在猪肺炎支原体。此外,该PCR系统用于分析来自育肥猪群的猪的鼻拭子样本。通过这种方法,我们能够在自然感染猪的鼻拭子中检测到猪肺炎支原体。然而,我们的结果表明,猪肺炎支原体仅在有限的时间段内可在鼻腔中检测到。

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