Lewin A, Freytag B, Meister B, Sharbati-Tehrani S, Schäfer H, Appel B (2003): Use of a quantitative TaqMan-PCR for the fast quantification of mycobacteria in broth culture, eukaryotic cell culture and tissue.
J. Vet. Med. B 50: 505-509.
The quantification of slow-growing mycobacteria such as Mycobacterium tuberculosis or M. bovis from in-vitro and in-vivo samples is complicated by their long generation time, their ability to form aggregates, and their capacity to persist in a state of dormancy. We compared different methods for the establishment of growth curves for broth cultures of M. bovis BCG. A quantitative TaqMan-PCR yielded results comparable to those obtained by protein quantification and measurement of the ATP content of the cultures. The quantitative TaqMan-PCR furthermore turned out to be particularly suitable for the measurement of multiplication of BCG within eukaryotic cells. Furthermore, it is a fast method allowing an estimation of the mycobacterial load in tissue long before colony counts can be obtained.