Location of MTBC primer-probes on the insertional elements, IS1081 and IS6110
The location of primer-probe sets for Mycobacterium tuberculosis complex insertional elements A) IS1081. B) IS6110. Primers designated “Miller” refer to Miller et al. 1997.
Primers and probes
Primers and probes used for real-time PCR identification of M. tuberculosis complex and M. bovis.
Initial performance of the primer and probe sets
The initial performance of the selected primer and probes designed to detect M. tuberculosis complex and M. bovis. A) Analytical sensitivity using M. bovis DNA. B) Calculated slope, R2 value, efficiency, and lowest detectable DNA concentration for each primer-probe set
Comparison of 3 DNA isolation methods using purification method A (magnetic bead) or method B (spin column)
Three DNA isolation methods, NaOH, phenol/chloroform and TE were evaluated using 3 replicates of 5 different samples. If all 3 replicates signaled positive, a mean and standard deviation of the Cτ were calculated. All 3 isolation methods were compared using 2 DNA purification methods; method A (magnetic bead) or method B (spin column).
Comparing DNA isolation and purification methods
Estimates and 95% confidence intervals comparing differences in means of two DNA isolation methods and two DNA purification methods. Using a linear model, the means differences in Cτ values per sample were generated and a 95% confidence interval calculated for each comparison. A result was considered statistically significant if the 95% CI did not span zero. A, B: Comparison of DNA Isolation methods using either DNA Purification method A or B. C, D: Comparison of DNA Purification Methods A and B using either DNA Isolation method 2 or 3.