PCRValidation of the real time PCR was done using known positiveand negative samples as well as the ATCC standard strain H37Rv spiked samples. Both the targets IS6110 as well as MPB64were detected in all dilutions in all the 3 repeat runs. Analyti-cal sensitivity of the real time multiplex PCR was found to beas low as 10 bacteria/mL of the sample. Cycling and meltingcurves of the multiplex PCR are shown in