Abstract:To set up the rapid technique for Mycobaeterium bovis by mycobacteriophage,the optimal test conditions were investigated for the detection of 16 non-Mtb strains of mycobacteria and other bacteria.It was demonstrated that the optimal working concentration of mycobacteriophage was 1×10~8 PFU/ml and the optimal condition for detection was the infection at 37℃ for 2 hours.Meanwhile,the concentration of virucidal agent to inactivate the mycobacteriophage completely was 100 mmol/L at room temperature for 10 min,and 4 mg/ml concentration of the logarithmic phase of mycobacteriophage was selected as the working concentration.Under these condition,the optimal results could be obtained,and the heat-inactivated or indicated bacteria were not infected with mycobacteriophage.It was found that positive results could be obtained by using this method of detection for control strain of M.bovis and M.smegmatis,but the negative results were demonstrated in 16 reference strains of non-Mtb strains of mycobacteria and 6 other bacteria.By using this method,60-120 CFU/ml of M.bovis could be detected.The intra-and inter-batch variation coeffiencies were all below 15%,as demonstrated by repeated testings,indicating a good repeatability.
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