Development of a biotinylated probe for the rapid detection of Mycoplasma gallisepticum

Mycoplasma gallisepticum causes respiratory diseases in the form of tracheitis and air sacculitis in chickens and turkeys. It is a major cause of reduced egg production, reduced hatchability, and downgrading of carcasses. Current means of diagnosis depend on the isolation and identification of the organisms, or on serological assays to detect serum antibodies. The evaluation of avian sera for M. gallisepticum antibodies is becoming more difficult to interpret, and thus less useful, due to the increasing use of killed M. gallisepticum vaccines. Maximum efficiency of M. gallisepticum disease control requires a rapid and sensitive identification system. A biotinylated total genome M. gallisepticum DNA probe was constructed by labeling the DNA with biotin-11-dUTP in a standard nick-translation reaction. Hybridization reactions with 100 ng/ml of biotinylated probe were capable of detecting 75 ng of M. gallisepticum target-DNA and 1.5 x 10(4) M. gallisepticum/ml within 24 h. The probe did hybridize to other mycoplasma DNAs, but to a greatly reduced degree.