牛分枝杆菌的噬菌体快速检测方法研究

建立牛分枝杆菌噬菌体快速检测技术,探讨最佳的检测条件,并将建立的方法用于牛分枝杆菌及16种常见非结核分枝杆菌和6种其他细菌的检测。结果表明,噬菌体工作浓度为1×108PFU/ml、37℃感染2h为最佳检测条件。杀毒剂浓度在100mmol/L,室温作用10min即可完全杀灭受试噬菌体。选择浓度为4mg/ml的对数生长期指示细菌为工作浓度,获得理想的检测结果。加热灭活的细菌和指示细菌不被噬菌体感染。牛分枝杆菌和耻垢分枝杆菌检测结果为阳性,16种常见非结核分枝杆菌和6种其他细菌检测结果均为阴性。该法可检测出60~120条/ml牛分枝杆菌。重复性试验表明,批内、批间变异系数均小于15%,重复性良好。

Rapid detection of Myeobaeterium bovis by mycobacteriophage

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.