Agence Française de Sécurité Sanitaire des Aliments (AFSSA), Unité d'Epidémiologie et de Bien-Etre du Porc, B.P. 53, 22440 Ploufragan, France.
Vet Microbiol. 2010 Jul 14;143(2-4):238-45. doi: 10.1016/j.vetmic.2009.12.001. Epub 2009 Dec 5.
Four sampling techniques for Mycoplasma hyopneumoniae detection, namely nasal swabbing, oral-pharyngeal brushing, tracheo-bronchial swabbing and tracheo-bronchial washing, were compared in naturally infected live pigs. In addition, a quantitative real-time PCR assay for M. hyopneumoniae quantification was validated with the same samples. 60 finishing pigs were randomly selected from a batch of contemporary pigs on a farm chronically affected by respiratory disorders. Each pig was submitted to nasal swabbing, oral-pharyngeal brushing, tracheo-bronchial swabbing and tracheo-bronchial washing. Nested-PCR and real-time PCR assays were performed on all samples. A Bayesian approach was used to analyze the nested-PCR results of the four sampling methods (i.e. positive or negative) to estimate the sensitivity and specificity of each method. M. hyopneumoniae was detected by nested-PCR in at least one sample from 70% of the pigs. The most sensitive sampling methods for detecting M. hyopneumoniae in live naturally infected pigs were tracheo-bronchial swabbing and tracheo-bronchial washing, as compared to oral-pharyngeal brushing and nasal swabbing. Swabbing the nasal cavities appeared to be the least sensitive method. Significantly higher amounts of M. hyopneumoniae DNA were found at the sites of tracheo-bronchial sampling than in the nasal cavities or at the oral-pharyngeal site (p<0.001). There was no difference between the tracheo-bronchial washing and the tracheo-bronchial swabbing results (p>0.05). Our study indicated that tracheo-bronchial swabbing associated with real-time PCR could be an accurate diagnostic tool for assessing infection dynamics in pig herds.
四种用于检测猪肺炎支原体的采样技术，即鼻拭子、口腔咽拭子、气管支气管拭子和气管支气管冲洗，在自然感染的活猪中进行了比较。此外，还使用相同的样本验证了猪肺炎支原体定量实时 PCR 检测方法。从一个长期受呼吸系统疾病影响的农场的一批当代猪中随机选择了 60 头育肥猪。每头猪都接受了鼻拭子、口腔咽拭子、气管支气管拭子和气管支气管冲洗。对所有样本进行了嵌套 PCR 和实时 PCR 检测。使用贝叶斯方法分析了四种采样方法（即阳性或阴性）的嵌套 PCR 结果，以估计每种方法的敏感性和特异性。在至少 70%的猪的至少一个样本中通过嵌套 PCR 检测到了猪肺炎支原体。与口腔咽拭子和鼻拭子相比，气管支气管拭子和气管支气管冲洗是检测活的自然感染猪中猪肺炎支原体最敏感的采样方法。鼻腔拭子似乎是最不敏感的方法。在气管支气管采样部位发现的猪肺炎支原体 DNA 量明显高于鼻腔或口腔咽部位（p<0.001）。气管支气管冲洗和气管支气管拭子的结果没有差异（p>0.05）。我们的研究表明，与实时 PCR 结合的气管支气管拭子可能是评估猪群感染动态的一种准确诊断工具。
