ABSTRACT
A recombinant phage library harbouring Mycoplasma meleagridis (MM) genomic DNA fragments was generated in the bacteriophage lambda gt11 expression vector. The library was screened for expression of MM specific antigens with a polyclonal antiserum that had been preadsorbed with antigens of the most common unrelated avian mycoplasma species. A 49-amino acid antigenic domain unique to MM was isolated, expressed in Escherichia coli, and its serodiagnostic potential was demonstrated. An antiserum raised against this MM-specific antigenic domain recognized a cluster of seven membrane-associated MM proteins with molecular masses ranging from 34 to 75 kDa. Overall, this study resulted in the identification of a potent serodiagnostic tool and revealed the complex antigenic nature of MM.
Subject(s)
Antigens, Bacterial/genetics , Cloning, Molecular , Mycoplasma meleagridis/genetics , Mycoplasma meleagridis/immunology , Animals , Antibodies, Bacterial/metabolism , Antigens, Bacterial/biosynthesis , Antigens, Bacterial/immunology , Base Sequence , DNA, Bacterial/chemistry , Escherichia coli/genetics , Immune Sera/metabolism , Immunoblotting/methods , Molecular Sequence Data , Mycoplasma Infections/diagnosis , Mycoplasma Infections/veterinary , Poultry Diseases/diagnosis , Poultry Diseases/microbiology , Species Specificity , TurkeysABSTRACT
We developed a duplex PCR assay targeting the hemagglutinin multigene families, vlhA and pMGA, of Mycoplasma synoviae and Mycoplasma gallisepticum, respectively. The assay proved to be specific and sensitive enough to justify its use for the simultaneous detection of the two major avian mycoplasma species from field isolates.
