I compared counting efficacies of CFU-enumerating method for the number of mycobacteria (
Mycobacterium intracellulare and
M. fortuitum) locating in cultured macrophages with that of microscopic counting method. Zymosan A-induced macrophages from ddY mice were infected with either
M. intracellulare or
M. fortuitum by incubation in 10% FBS-RPMI 1640 medium containing the organisms for 1 hr, thereafter thoroughly washed to remove extracellular bacilli, and cultured for 3 to 5 days. At intervals, macrophages were thoroughly rinsed and subjected to either CFU-enumeration or microscopic counting, as follows. In the former method, macrohages were lysed with 0.2% Tween 80-distilled water by sonication using Handy Sonic and CFUs were counted on 7H11 agar plates. In the latter method, the number of acid-fast bacilli was counted by microscopy for macrophages after Ziehl-Toda's staining. The number of bacteria by the CFU-enumerating method was much greater than that by the microscopic counting method.
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