Development of an ELISA using a recombinant P46-like lipoprotein for diagnosis of Mycoplasma pulmonis infection in rodents.
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.
Laboratory of Veterinary Biochemistry, School of Veterinary Medicine, Faculty of Agriculture, Tottori University, Tottori 680-8553, Japan.
Atsushi
A
Asano
Daisuke
D
Torigoe
Nobuya
N
Sasaki
Takashi
T
Agui
D000595:Amino Acid Sequence; D000818:Animals; D000830:Animals, Laboratory / Q000382:microbiology*; D001426:Bacterial Proteins / Q000235:genetics; D003001:Cloning, Molecular; D017931:DNA Primers / Q000235:genetics; D018076:DNA, Complementary / Q000235:genetics; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods / Q000662:veterinary*; D004926:Escherichia coli; D005982:Glutathione Transferase; D015151:Immunoblotting / Q000662:veterinary; D051379:Mice; D008810:Mice, Inbred C57BL; D008969:Molecular Sequence Data; D009175:Mycoplasma Infections / Q000175:diagnosis / Q000662:veterinary*; D045803:Mycoplasma pulmonis / Q000235:genetics*; D051381:Rats; D012376:Rodent Diseases / Q000175:diagnosis* / Q000382:microbiology*; D016415:Sequence Alignment
D001426:Bacterial Proteins; D017931:DNA Primers; D018076:DNA, Complementary; C093625:surface antigen p46, Mycoplasma; D005982:Glutathione Transferase