RESUMO
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.
Assuntos
Antígenos de Bactérias/genética , Clonagem Molecular , Mycoplasma meleagridis/genética , Mycoplasma meleagridis/imunologia , Animais , Anticorpos Antibacterianos/metabolismo , Antígenos de Bactérias/biossíntese , Antígenos de Bactérias/imunologia , Sequência de Bases , DNA Bacteriano/química , Escherichia coli/genética , Soros Imunes/metabolismo , Immunoblotting/métodos , Dados de Sequência Molecular , Infecções por Mycoplasma/diagnóstico , Infecções por Mycoplasma/veterinária , Doenças das Aves Domésticas/diagnóstico , Doenças das Aves Domésticas/microbiologia , Especificidade da Espécie , PerusRESUMO
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.
