Comparative study of two in vitro dormant models for Mycobacterium tuberculosis
JIANG Li-sha1, 2, YAO Yi-yong1, ZHANG Li1, LIU Yan1, GUO Shu-liang1
1. Department of Respiratory and Critical Care Medicine,the First Affiliated Hospital of Chongqing Medical University,Chongqing 400016,China; 2. The first branch the First Affiliated Hospital of Chongqing Medical University,Chongqing 400016,China
Abstract：To compare the oxygen-deficit model with the multiple-stress model,and understand their pros and cons,the oxygen-deficit model was constructed by applying gradually low oxygen. The multiple-stress model was developed by applying combined stress of low oxygen (5% O2),high carbon dioxide (10% CO2),faintly acid (pH 5.0) and lack of nutrient (10% 7H9 broth). After both the models were constructed for 20 days and 30 days,respectively,the sample was collected. Loss of acid fastness and accumulation of lipid bodies was observed through confocal microscopy after dual staining of Mycobacterium tuberculosis (Mtb) with the combination Auramine-O and Nile Red. Cell wall thickness of Mtb was observed by electron microscope. And the drug-resistance rate to rifampicin (Rif) was also detected. At 30th day,equivalent Mtb of two models was subcultured into fresh 7H9 broth to observe the auto-resuscitation. We discovered that Mtb of two models showed loss of acid fastness and accumulation of lipid bodies but no transformation of cell wall at 20th day. Mtb of multiple-stress model almost showed loss of acid fastness and accumulation of lipid bodies at 30th day. However,portions of Mtb did not show loss of acid fastness and accumulation of lipid bodies in oxygen-deficit model. At 30th day,cell wall thickness of Mtb was observed in multiple-stress model but did not in oxygen-deficit model. The drug-resistance rate to rifampicin of multiple-stress model was obviously higher than that of oxygen-deficit model,and the amount of Mtb in multiple-stress model was smaller than that of oxygen-deficit model after auto-resuscitation in fresh 7H9 broth. In Conclusion,Mtb could develope dormant state more rapid in multiple-stress model,and its dormant state was more stable.
江莉莎, 姚义勇, 张莉, 柳岩, 郭述良. 缺氧模型和多因素模型构建结核休眠菌模型比较研究[J]. 中国人兽共患病学报, 2016, 32(4): 327-331.
JIANG Li-sha, YAO Yi-yong, ZHANG Li, LIU Yan, GUO Shu-liang. Comparative study of two in vitro dormant models for Mycobacterium tuberculosis. Chinese Journal of Zoonoses, 2016, 32(4): 327-331.
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2016, Vol. 32 
