噬菌体Guo1复苏结核休眠菌的初步研究
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国家自然科学基金面上项目(81570010);国家临床重点专科专项(2012-649)


Resuscitation of mycobacteriophage Guo1 in dormant cells of M.tuberculosis
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    摘要:

    目的:探索分枝杆菌噬菌体Guo1对结核休眠菌的复苏作用。方法:采用缺钾Sauton培养基培养对数生长期结核菌构建结核休眠菌模型;采用药敏检测、透射电子显微镜(电镜)观察和金胺O-尼罗红双荧光染色作为模型检测方法;采用菌落计数作为复苏的检测指标。结果:缺钾结核休眠菌模型构建成功。混合培养第1天时,空白对照组、上清组、TM4组、Guo1组菌落计数分别为(2.67±0.58)×102、(3.00±1.00)×102、(2.67±0.58)×102、(2.33±0.58)×102 CFU/mL;培养第15天时,各组菌落计数分别为(7.33±1.53)×104、(6.67±2.08)×1010、(3.00±1.00)×1010、(3.33±1.53)×1010CFU/mL。结论:分枝杆菌噬菌体Guo1能够复苏结核休眠菌。

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    Objective:To investigate the resuscitative potential of mycobacteriophage Guo1 on dormant cells of M.tuberculosis.Methods:Dormant cells of M.tuberculosis were induced by potassium-deficient culture of logarithmic phase of M.tuberculosis at 37 ℃,which were proven through drug sensitivity test,transmission electron microscope(TEM) and auramine O-Nile red double fluorescent staining. M.tuberculosis colony count was used to detect whether the dormant cells of M.tuberculosis had been resuscitated by Guo1. Results:Dormant cells of M.tuberculosis were obtained successfully by potassium-deficient cultivation for 30 days at 37 ℃. The concentration of M.tuberculosis in control,supernatant(SN),TM4 and Guo1 groups was (2.67±0.58)×102,(3.00±1.00)×102,(2.67±0.58)×102,(2.33±0.58)×102 CFU/mL,respectively,after culture for 1 d. And the concentration of M.tuberculosis in above groups increased to(7.33±1.53)×104,(6.67±2.08)×1010,(3.00±1.00)×1010,(3.33±1.53)×1010 CFU/mL at the 15th day,respectively. Conclusion:Mycobacteriophage Guo1 can resuscitate dormant cells of M.tuberculosis.

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杨 婷,杜丽娟,曹 俊,郭述良.噬菌体Guo1复苏结核休眠菌的初步研究[J].南京医科大学学报(自然科学版),2017,(10):1234-1238

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  • 收稿日期:2016-11-06
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  • 在线发布日期: 2017-11-06
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