Asano Atsushi, Torigoe Daisuke, Sasaki Nobuya, Agui Takashi
Laboratory of Veterinary Biochemistry, School of Veterinary Medicine, Faculty of Agriculture, Tottori University, Tottori 680-8553, Japan.
J Vet Med Sci. 2014 Mar 1;76(2):151-7. doi: 10.1292/jvms.13-0308. Epub 2013 Sep 20.
Mycoplasma pulmonis is one of the most prevalent bacterial pathogens that infects laboratory mice and rats. To develop an M. pulmonis-specific antigen for serological diagnosis, we cloned the cDNA of P46-like lipoprotein (P46L), an M. pulmonis putative periplasmic protein. P46L is a homolog of P46, an M. hyopneumoniae antigen. We produced recombinant P46L fused to glutathione S-transferase (GST) in Escherichia coli. Immunoblot analysis revealed that sera from Mycoplasma-infected mice and rats contained anti-P46L antibodies. We developed an ELISA using the recombinant P46L-GST protein as an antigen. Thirteen of the 14 samples from rats naturally infected with M. pulmonis were determined to be positive according to the commercial ELISA (MONILISA Myco) and positive by our ELISA. Furthermore, 18/19 samples from mice experimentally infected with M. pulmonis were positive using our P46L-GST ELISA. In contrast, only 8/19 samples from infected mice were positive by the commercial ELISA. Our results indicate that P46L-GST was an appropriate antigen for developing a serological test to determine M. pulmonis infection in laboratory mice and rats.
肺支原体是感染实验小鼠和大鼠的最常见细菌病原体之一。为了开发一种用于血清学诊断的肺支原体特异性抗原，我们克隆了P46样脂蛋白（P46L）的cDNA，P46L是一种肺支原体假定的周质蛋白。P46L是猪肺炎支原体抗原P46的同源物。我们在大肠杆菌中生产了与谷胱甘肽S-转移酶（GST）融合的重组P46L。免疫印迹分析显示，来自感染支原体的小鼠和大鼠的血清中含有抗P46L抗体。我们使用重组P46L-GST蛋白作为抗原开发了一种ELISA。根据商业ELISA（MONILISA Myco），14份自然感染肺支原体的大鼠样本中有13份被判定为阳性，并且通过我们的ELISA也呈阳性。此外，使用我们的P46L-GST ELISA，19份实验感染肺支原体的小鼠样本中有18份呈阳性。相比之下，商业ELISA检测感染小鼠的样本中只有8/19呈阳性。我们的结果表明，P46L-GST是开发用于确定实验小鼠和大鼠肺支原体感染的血清学检测的合适抗原。
