Sato N, Fieldsteel A H
J Clin Microbiol. 1977 Mar;5(3):326-8. doi: 10.1128/jcm.5.3.326-328.1977.
A new method of enumerating Mycobacterium leprae has been developed. Suspensions containing the organisms were filtered through a polycarbonate membrane filter (25-mm diameter, 0.4-micronm pore size, 10-micronm thick; Nucleopore) to concentrate the organisms. The membrane was then mounted on a glass slide and stained with a standard acid-fast stain. Finally, the membrane was treated with a small amount of chloroform to fix it to the slide and make it transparent. This method enabled us to detect M. leprae in quantities as small as 4.98 X 10(2) regardless of the total volume of the original material. Comparison with a standard method for enumerating M. leprae showed that both methods gave similar results when the organisms counted by the standard method were present in sufficient quantity for reproducibility. Because the least number of organisms that can be detected with the standard method is 10(4) ml and because the organisms detected with the new method could be concentrated on the polycarbonate filter from a large amount of infected fluid, a substantial number of suspensions were shown by the new method, but not by the standard method, to contain M. leprae.
一种新的麻风分枝杆菌计数方法已被开发出来。含有该微生物的悬浮液通过聚碳酸酯膜过滤器（直径25毫米，孔径0.4微米，厚度10微米；核孔滤膜）进行过滤，以浓缩微生物。然后将滤膜安装在载玻片上，用标准抗酸染色法进行染色。最后，用少量氯仿处理滤膜，使其固定在载玻片上并变得透明。这种方法使我们能够检测到低至4.98×10²数量的麻风分枝杆菌，而不管原始材料的总体积如何。与麻风分枝杆菌计数的标准方法进行比较表明，当标准方法计数的微生物数量足够用于重复性时，两种方法给出的结果相似。由于标准方法能够检测到的最少微生物数量为10⁴/ml，并且由于用新方法检测到的微生物可以从大量感染液中浓缩在聚碳酸酯滤膜上，新方法显示出大量悬浮液含有麻风分枝杆菌，而标准方法却未检测到。
