DEVELOPMENT OF A DNA PROBE FOR THE RAPID DETECTION OF OF MYCOMPLASMA GALLISEPTICUM

THE OBJECTIVE OF THIS PROJECT IS TO CONSTRUCT A MYCOMPLASMA GALLISEPTICUM BIOTINYLATED DNA PROBE. ECONOMIC LOSSES FROM M. GALLISEPTICUM INFECTION ARE ESTIMATED TO BE $118 MILLION ANNUALLY IN THE UNITED STATES. MINIMUM EFFICIENCY OF M. GALLISEPTICUM DISEASE CONTROL REQUIRES A RAPID AND SENSITIVEIDENTIFICATION SYSTEM SUCH AS A PROBE. MYCOPLASMA GAL GALLISEPTICUM DNA WILL BE LABELED WILL BIOTIN-11-DUTP IN A STANDARD NICK-TRANSLATION REACTION. TARGET DNA HYBRIDIZED WITH THE BIOTINYLATED PROBE ON A NITROCELLULOSE FILTER WILL BE INCUBATED WITH STREPTAVIDIN AND POLYALKALINE PHOSPHASTASE. UPON ADDITION OF AN ENZYME/SUBSTRATE COMPLEX, THE HYBRIDIZED DNA WILL APPEAR AS A BLUE SPOT. RESULTS CAN BE OBTAINED IN 24-36 HOURS, WHICH IS CONSIDERABLY FASTER THAN ROUTINE METHODS CURRENTLY IN USE.