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Mycoplasma synoviae surface protein MSPB as a recombinant antigen in an indirect ELISA

Amir H. Noormohammadi

School of Veterinary Science, The University of Melbourne, Parkville, Victoria, Australia 3052

Present address: WEHI, Post Office, Royal Melbourne Hospital, VIC 3050, Australia.

Author for correspondence: Amir H. Noormohammadi. Tel: + 61 3 9345 2475. Fax: + 61 3 9347 0852. e-mail: amir-h@wehi.edu.au

ABSTRACT

Mycoplasma synoviae is a poultry pathogen causing respiratory disease and synovitis. A number of serological assays have been developed for diagnosis of M. synoviae infection; however, they lack sensitivity and/or are prone to false-positive reactions. Using a combination of PCR and expression cloning, four overlapping regions (regions 1-4) of the surface antigen MSPB of M. synoviae WVU-1853 were expressed in a bacterial expression system.

Immunostaining of the resultant polypeptides with chicken sera raised against different M. synoviae strains, or Mycoplasma gallisepticum S6, suggested that region 4 contained a highly antigenic and species-specific domain (amino acids 178-213) of MSPB. A fusion protein of region 4 was expressed in the pMAL expression system and purified from cold-osmotic-shock fluids of Escherichia coli cells for use in an indirect ELISA. The potential of the purified antigen for detection of M. synoviae antibodies was assessed with sera obtained from chickens experimentally infected with different strains of M. synoviae or M. gallisepticum, or from uninoculated chickens. All the sera from M. synoviae-inoculated chickens provided higher absorbance values than those from M. gallisepticum-inoculated or uninoculated chickens. Chickens inoculated with M. synoviae 86079/7NS had detectable increases of serum anti-MSPB immunoglobulins at day 7 after inoculation. These studies have identified the most antigenic region of one of the major species-specific surface proteins of M. synoviae, and shown the potential of this protein as a serodiagnostic reagent.

The GenBank accession number for the vlhA gene sequence is AF035624.

Abbreviations: HRP, horseradish peroxidase; RSA, rapid serum agglutination; RU, relative units; SPF, specific-pathogen-free.

Received 2 February 1999; revised 16 April 1999; accepted 20 April 1999.

This Article Abstract Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via CrossRef Citing Articles via Google Scholar Google Scholar Articles by Noormohammadi, A. H. Articles by Browning, G. F. Search for Related Content PubMed PubMed Citation Articles by Noormohammadi, A. H. Articles by Browning, G. F. Agricola Articles by Noormohammadi, A. H. Articles by Browning, G. F.