Use of a quantitative TaqMan-PCR for the fast quantification of mycobacteria in broth culture, eukaryotic cell culture and tissue.
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 bacille Calmette-Guérin (BCG). A quantitative TaqMan-PCR yielded results comparable with 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.
Lewin
A
A
Robert Koch-Institut, Nordufer 20, 13353 Berlin. lewina@rki.de
Freytag
B
B
Meister
B
B
Sharbati-Tehrani
S
S
Schäfer
H
H
Appel
B
B
D000818:Animals; D002478:Cells, Cultured / Q000502:physiology; D003470:Culture Media; D004269:DNA, Bacterial / Q000032:analysis; D005260:Female; D006168:Guinea Pigs; D008264:Macrophages / Q000382:microbiology; D008297:Male; D009163:Mycobacterium bovis / Q000235:genetics / Q000502:physiology*; D009169:Mycobacterium tuberculosis / Q000235:genetics / Q000502:physiology*; D016133:Polymerase Chain Reaction / Q000662:veterinary*; D011237:Predictive Value of Tests; D013234:Stem Cells
D003470:Culture Media; D004269:DNA, Bacterial