Satish Rajitha, Desouza Anita
Department of Microbiology, SIES College of Arts, Science and Commerce, Affiliated to Mumbai University, Mumbai, Maharashtra, India.
Int J Mycobacteriol. 2019 Apr-Jun;8(2):170-174. doi: 10.4103/ijmy.ijmy_42_19.
Mycobacteriophages are viruses that infect Mycobacterium spp. Till date, 10427 mycobacteriophages have been isolated and 1670 mycobacteriophage genomes have been sequenced https://phagesdb.org/hosts/genera/1/ (cited on 30th December,2018). In the previous study, 10 different mycobacteriophages from 14 soil samples were isolated, by qualitative plaque formation method using Mycobacterium smegmatis as host. Among these, three phages were found to infect four different species of Mycobacterium, i.e., Mycobacterium fortuitum subsp. fortuitum MTCC993, Mycobacterium kansasii MTCC3058, Mycobacterium avium subsp. avium MTCC1723, and Mycobacterium tuberculosis MTCC300, besides the host M. smegmatis. The phage lysates were concentrated by polyethylene glycol (PEG) precipitation. One of the three phages showing host diversity was selected for further study. The various phage growth parameters such as incubation temperature, time of adsorption, host cell density and effect of cations were standardised.
The studies were done by qualitative and quantitative plaque assay method.
The phage selected for further study showed an optimum adsorption time of 15 min. The optimum temperature for propagation was found to be 37°C. The phage was found to be stable at 42°C. In the presence of calcium, the phage showed a higher rate of infectivity.
Understanding the biology of mycobacteriophages and their host diversity is the key to understanding mycobacterial systems. This could be the first step toward exploiting the potential of phages as therapeutic agents.
分枝杆菌噬菌体是感染分枝杆菌属细菌的病毒。截至目前,已分离出10427种分枝杆菌噬菌体,1670种分枝杆菌噬菌体基因组已完成测序(https://phagesdb.org/hosts/genera/1/,引用日期为2018年12月30日)。在之前的研究中,通过以耻垢分枝杆菌为宿主的定性噬菌斑形成方法,从14份土壤样本中分离出10种不同的分枝杆菌噬菌体。其中,除了宿主耻垢分枝杆菌外,发现三种噬菌体可感染四种不同的分枝杆菌,即偶然分枝杆菌偶然亚种MTCC993、堪萨斯分枝杆菌MTCC3058、鸟分枝杆菌鸟亚种MTCC1723和结核分枝杆菌MTCC300。噬菌体裂解物通过聚乙二醇(PEG)沉淀进行浓缩。选择三种表现出宿主多样性的噬菌体之一进行进一步研究。对各种噬菌体生长参数,如培养温度、吸附时间、宿主细胞密度和阳离子的影响进行了标准化。
通过定性和定量噬菌斑测定法进行研究。
选择进行进一步研究的噬菌体的最佳吸附时间为15分钟。发现其增殖的最佳温度为37°C。该噬菌体在42°C时稳定。在钙存在的情况下,噬菌体显示出更高的感染率。
了解分枝杆菌噬菌体的生物学特性及其宿主多样性是理解分枝杆菌系统的关键。这可能是开发噬菌体作为治疗剂潜力的第一步。