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Development of an ELISA using a recombinant P46-like lipoprotein for diagnosis of Mycoplasma pulmonis infection in rodents. | LitMetric

Development of an ELISA using a recombinant P46-like lipoprotein for diagnosis of Mycoplasma pulmonis infection in rodents.

J Vet Med Sci

Laboratory of Veterinary Biochemistry, School of Veterinary Medicine, Faculty of Agriculture, Tottori University, Tottori 680-8553, Japan.

Published: March 2014

AI Article Synopsis

  • Mycoplasma pulmonis is a common bacterial pathogen affecting lab mice and rats, and researchers aimed to create a specific antigen for diagnosing infections.
  • They successfully cloned the cDNA of a protein called P46-like lipoprotein (P46L) and produced it in E. coli, confirming that infected rodents had antibodies against this protein.
  • Their newly developed ELISA test, using the P46L-GST protein, outperformed a commercial test, detecting M. pulmonis infection more effectively in both rats and mice.

Article Abstract

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.

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Source
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3982827PMC
http://dx.doi.org/10.1292/jvms.13-0308DOI Listing

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