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The Critical Role of DNA Extraction for Detection of Mycobacteria in Tissues

Figure 4

in vitro limit of quantification (on the left) estimated by reproducible TaqMan real-time PCR amplifications of MAP0865 (n=63) and ext-RD9 (n=21) for MAP K-10 (A) and M. tuberculosis H37Rv quantification (B), and limits of quantification in tissues (on the right) estimated by distribution analyses of sample number according to MAP K-10 (A) and M. tuberculosis H37Rv (B) densities, estimated by TaqMan real-time PCR targeting MAP0865 (n=40 spleen, n=40 liver) or ext-RD9 (n=7 spleen, n=9 lung), respectively.

All these accurate real-time PCR results are observed for a host DNA amount < 3 µg in a final volume of 25 µl, and DNA purity at 260/280 nm ratio = 1.89±0.08.

Figure 4