Ryan C, Nguyen B T, Sullivan S J
One Cell Systems, Inc., Cambridge, Massachusetts 02139, USA.
J Clin Microbiol. 1995 Jul;33(7):1720-6. doi: 10.1128/jcm.33.7.1720-1726.1995.
Effective control of tuberculosis transmission in vulnerable population groups is dependent on rapid identification of the infectious agent and its drug susceptibility. However, the slow growth rate of mycobacteria has undermined the ability to quickly identify antimicrobial resistance. These studies describe a mycobacterial growth assay based on microencapsulation technology used in conjunction with flow cytometric analysis. Mycobacteria were encapsulated in agarose gel microdrops approximately 25 microns in diameter, and colony growth was monitored by using flow cytometry to evaluate the intensity of auramine staining after culture for various times at 37 degrees C. By this method, colony growth of Mycobacterium bovis and M. smegmatis could be quantified within 1 to 3 days after encapsulation. Inhibition of growth by rifampin and isoniazid was also evaluated in this time period, and the presence of an isoniazid-resistant subpopulation representing 3% of the total microorganisms could be detected. This use of encapsulation and flow cytometry has the potential to facilitate rapid and automated evaluation of inhibition of growth by antimicrobial agents and shorten the time frame for analysis of clinical specimens.
在脆弱人群中有效控制结核病传播取决于快速鉴定感染病原体及其药敏情况。然而，分枝杆菌生长速度缓慢削弱了快速鉴定抗菌药物耐药性的能力。这些研究描述了一种基于微囊化技术并结合流式细胞术分析的分枝杆菌生长测定法。分枝杆菌被包裹在直径约25微米的琼脂糖凝胶微滴中，通过在37℃培养不同时间后使用流式细胞术评估金胺染色强度来监测菌落生长。通过这种方法，牛分枝杆菌和耻垢分枝杆菌的菌落在包裹后1至3天内即可进行定量。在此时间段内还评估了利福平和异烟肼对生长的抑制作用，并且可以检测到占总微生物3%的异烟肼耐药亚群的存在。这种微囊化和流式细胞术的应用有可能促进对抗菌药物生长抑制的快速和自动化评估，并缩短临床标本分析的时间框架。
