In situ hybridisation for identification and differentiation of Mycoplasma hyopneumoniae, Mycoplasma hyosynoviae and Mycoplasma hyorhinis in formalin-fixed porcine tissue sections
In situ hybridisation for identification and differentiation of Mycoplasma hyopneumoniae, Mycoplasma hyosynoviae and Mycoplasma hyorhinis in formalin-fixed porcine tissue sections
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DOI：
10.1034/j.1600-0463.2001.d01-129.x
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发表时间：
2001-10-01
期刊：
影响因子：
2.8
通讯作者：
Friis, NF
中科院分区：
文献类型：
--
作者：
Boye, M;Jensen, TK;Friis, NF
设计了靶向16S核糖体RNA的寡核苷酸探针，用于使用荧光原位杂交法鉴定猪支原体猪支原体、猪鼻支原体和猪滑液支原体。使用纯培养物以及人工存在支原体的猪组织切片评价探针的特异性，发现探针对靶生物具有特异性。该试验应用于28份组织样本的切片，这些组织样本来自通过培养确定感染三种支原体中的一种或多种的猪。M.和M.根据肺切片的培养，对9头患有卡他至脓性支气管肺炎的猪的猪鼻进行了鉴定。同样，在8例化脓性心包炎中，M。hypopalanum，M. hyorhinis和M.猪滑液绦虫是感染因子，根据培养，并通过原位杂交正确鉴定。在I I关节培养阳性的M。在猪滑液囊尾蚴中，探针只能识别M.在8例病例中，可能是由于组织切片中微生物数量较少所致。原位杂交技术不仅适用于诊断和实验研究，而且也可作为致病机制研究的工具。
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 I I 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 toot for pathogenesis studies.