猪肺炎支原体气溶胶富集检测技术的建立(英文)

Establishment of a Collection and Detection Technology of Mycoplasma hyopneumoniae in Aerosol

[目的]建立一个高效的猪肺炎支原体气溶胶富集和检测技术,以弥补临床上猪肺炎支原体气溶胶检测技术的空白。[方法]结合液体冲击式采样器和过滤式采样器的原理,自行设计一个气溶胶双重富集装置,并在密闭环境下采集人工制备的猪肺炎支原体气溶胶,荧光定量PCR检测以确定该装置的采集效率。再通过收集不同猪群中的气溶胶进行检测以确定该装置临床应用的可行性。首先,收集人工感染Mhp阴性仔猪后不同时间猪舍内的气溶胶;其次,收集1个猪支原体肺炎发病猪场和1个阳性未发病猪场的产房、保育舍和育肥舍共11个猪舍的气溶胶,荧光定量PCR或套式PCR检测所收集的气溶胶。[结果]实验室模拟采样的采集效率达到(37.04±6.43)%,人工感染猪肺炎支原体后7天即在饲养房间的气溶胶中检到猪肺炎支原体;2个猪场共11个不同猪舍气溶胶中猪肺炎支原体检测结果均为阳性,检出率100%。[结论]成功建立了一个高敏感性的猪肺炎支原体气溶胶富集检测技术,适用于临床上猪肺炎支原体气溶胶的检测。

ObjectiveThis study was to establish a simple method for collecting and detecting Mycoplasma hyopneumoniae （Mhp） in aerosol. MethodBased on the mechanisms of liquid impinger and filtration sampler, a double concentration aerosol sampler was designed for collecting Mhp aerosol. Firstly, the collection was performed in a closed environment full of artificial aerosol of Mhp. Secondly, collection efficiency was detected by real-time PCR. Thereafter, the clinical feasibility of the designed equipment was tested by collecting aerosol samples in different pig herds. In one assay, the samples were collected at different times from one pig house challenged with Mhp. In another assay, the samples was collected from the delivery room, nursery and fattenning house of a MPS outbreak farm as well as a Mhp infection positive pig farm without obvious clinical symptoms. All the aerosol samples were then detected by real-time PCR or nested PCR. ResultThe collection efficiency of the designed bioaerosol sampler was （37.04±6.43） %, Mhp could be detected 7 d after intratracheal challenge with pneumonic lung homogenate suspension. Aerosol samples of 11 pig houses from the two Mhp positive pig farms with or without clinical symptoms all showed a positive result of PCR, the positivity rate was 100%. ConclusionA high sensitive collecting and detecting technology of aerosol was successfully established, which can be applied to clinical detection of Mhp in aerosol.