Boye M, Jensen T K, Ahrens P, Hagedorn-Olsen T, Friis N F
Danish Veterinary Laboratory, Copenhagen, Denmark.
APMIS. 2001 Oct;109(10):656-64. doi: 10.1034/j.1600-0463.2001.d01-129.x.
Oligonucleotide probes targeting 16S ribosomal RNA were designed for species-specific identification of the porcine mycoplasmas Mycoplasma hyopneumoniae, Mycoplasma hyorhinis and Mycoplasma hyosynoviae using a fluorescent in situ hybridisation assay. The specificity of the probes was evaluated using pure cultures as well as porcine tissue sections with artificial presence of mycoplasma, and the probes were found specific for the target organisms. The assay was applied on sections of 28 tissue samples from pigs infected with one or more of the three Mycoplasma species as determined by cultivation. M. hyopneumoniae and M. hyorhinis were identified in accordance with cultivation in lung sections from nine pigs affected by catarrhal to purulent bronchopneumonia. Likewise, in eight cases of fibrinous pericarditis, M. hyopneumoniae, M. hyorhinis and M. hyosynoviae were the infectious agents according to cultivation and were correctly identified by in situ hybridisation. Out of 11 joints cultivation positive for M. hyosynoviae, the probe was only able to identify M. hyosynoviae in eight cases probably due to a low number of microorganisms in the tissue sections. The in situ hybridisation assay is well suited for use in diagnostic and experimental work as well as a tool for pathogenesis studies.
设计了靶向16S核糖体RNA的寡核苷酸探针，用于通过荧光原位杂交试验对猪肺炎支原体、猪鼻支原体和猪滑膜支原体进行种特异性鉴定。使用纯培养物以及人工存在支原体的猪组织切片评估探针的特异性，发现这些探针对目标生物体具有特异性。该试验应用于来自28个组织样本的切片，这些样本来自经培养确定感染了三种支原体中的一种或多种的猪。在9头患有卡他性至脓性支气管肺炎的猪的肺切片中，根据培养结果鉴定出猪肺炎支原体和猪鼻支原体。同样，在8例纤维素性心包炎病例中，根据培养结果，猪肺炎支原体、猪鼻支原体和猪滑膜支原体是感染病原体，并通过原位杂交得到正确鉴定。在11个关节培养对猪滑膜支原体呈阳性的样本中，该探针仅在8例中能够鉴定出猪滑膜支原体，这可能是由于组织切片中微生物数量较少。原位杂交试验非常适合用于诊断和实验工作以及作为发病机制研究的工具。
