Stemke G W
Department of Microbiology, University of Alberta, Edmonton, Canada.
Mol Cell Probes. 1989 Sep;3(3):225-32. doi: 10.1016/0890-8508(89)90003-0.
Enzootic porcine pneumonia is caused by Mycoplasma hyopneumoniae. Since the disease is of world-wide importance it is important to detect and identify the causative agent. In experience laboratories this mycoplasma can usually be detected by culture but its identification still is difficult and time consuming. We have cloned random Eco R1 fragments of M. hyopneumoniae DNA to M13mp19 and used the resultant recombinant to produce a probe capable of detecting approximately 10 pg of the mycoplasma DNA (10(4) organisms). By using appropriate stringency the test was made specific for M. hyopneumoniae, although at lower stringency reaction was positive with Mycoplasma flocculare at 1000 x the concentration limit. The assay did not detect M. hyopneumoniae in DNA from lungs of chronically infected animals but it did react with DNA isolated from the organisms cultured from the infected lung material.
地方性猪肺炎由猪肺炎支原体引起。由于该疾病在全球范围内具有重要意义,因此检测和鉴定病原体至关重要。在有经验的实验室中,这种支原体通常可以通过培养检测到,但其鉴定仍然困难且耗时。我们已将猪肺炎支原体DNA的随机Eco R1片段克隆到M13mp19中,并使用所得重组体产生一种能够检测约10 pg支原体DNA(10⁴个生物体)的探针。通过使用适当的严谨性,该测试对猪肺炎支原体具有特异性,尽管在较低严谨性下,浓度极限为1000倍时与絮状支原体的反应呈阳性。该检测方法未在慢性感染动物肺部的DNA中检测到猪肺炎支原体,但它确实与从感染肺组织培养的生物体中分离的DNA发生反应。