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Lysis effect of phage Guo1 and Leo on Mycobacterium tuberculosis in the dormant stage of different models |
JIANG Lisha1,2 GUO Shuliang1 |
1.Department of Respiratory and Critical Care Medicine, the First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, China;
2.Department of Respiratory Medicine, the First Branch, the First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, China |
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Abstract Objective To investigate the lysis effect of phage Guo1 and Leo on Mycobacterium tuberculosis in dormant stage. Methods ClustalX software was used for multiple sequence comparison. Motif3 motifs of phage Guo1 and Leo were searched. The model of Mycobacterium tuberculosis in dormant stage was constructed by hypoxia model and potassium deficiency model respectively. Hypoxic and potassium deficiency mode induce domant TB bacterial fluid, they were respectively randomly divided into 5 groups, each group was set with 3 duplicate tubes, and each tube was filled with 2 mL bacterial liquid. Then 100 liters of Isoniazid (final concentration: 5 g/mL, Isoniazid group), Rifampicin (final concentration: 5 g/mL, Rifampicin group), phage Leo (titer: 2.6×108 PFU/mL, Leo group), and phage Guo1 (titer: 3.26×108 PFU/mL, Guo1 group) were added to the test tubes of each group, and the control group was added with the same amount of ddH2O. The drug resistance to Isoniazid and Rifampicin in dormant tuberculosis bacteria and the lysis of phage in dormant Tuberculosis bacteria were detected by most probable number method. Results The motif3 motif was found in the tape rule proteins of phages Guo1 and Leo, and the motif3 motif contained 6 highly conserved amino acid residues and 1 highly conserved isoleucine-tryptophan site. When used as hypoxia model for dormant Tuberculosis bacteria, compared with Rifampin group and Isoniazid group, the bacteria quantity of Guo1 group decreased significantly (P < 0.05). Compared with Rifampicin group, the amount of bacteria in Leo group decreased significantly (P < 0.05). The drug resistance rate of potassium deficiency model to Isoniazid and Rifampicin was significantly higher than that of hypoxia model. Conclusion Compared with hypoxia model, potassium deficiency model can induce Tuberculosis bacterium to enter deeper dormant state. Phage Guo1 has cleavage effect on dormant tuberculosis bacteria induced by hypoxia model, and phage Leo has cleavage effect on dormant Tuberculosis bacteria induced by potassium deficiency model.
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