分枝杆菌噬菌体在云南省的首次分离鉴定和特性研究

doi:10.19982/j.issn.1000-6621.20250248

摘要/Abstract

摘要：

目的: 自云南省分离分枝杆菌噬菌体,并阐明其表型及基因组学特性。方法: 在云南省昆明市、大理州、怒江州、德宏州、玉溪市、临沧市、普洱市、昭通市、保山市,采集医院污水样、土样和粪便样等标本。以耻垢分枝杆菌mc²155为宿主菌,采用直接法和富集法从样本中分离分枝杆菌噬菌体;对分离的分枝杆菌噬菌体进行生物学特性、全基因组序列测定及基因组分析。结果: 从186份样本中,共分离到3株分枝杆菌噬菌体,选择其中1株(命名为YN1)进行研究。研究表明,YN1最佳感染复数为0.1,生长潜伏期约为330min,环境因素如温度、酸碱度、紫外线和乙醇对YN1有一定影响。基因组分析显示,YN1属线性双链DNA病毒;比较基因组学表明,YN1在分类学上属分枝杆菌Ⅰ簇噬菌体。结论: 本研究首次从云南省内分离出分枝杆菌噬菌体,经鉴定和特性研究,YN1为1株新的分枝杆菌Ⅰ簇噬菌体,丰富了国内分枝杆菌噬菌体库,并为分枝杆菌噬菌体的进一步开发和利用提供基础。

关键词: 分枝杆菌噬菌体, 生物学, 基因组, 表型, 云南省

Abstract:

Objective: To isolate Mycobacteriophages from Yunnan Province and elucidate their phenotypic and genomic characteristics. Methods: Hospital sewage samples, soil samples, and fecal samples were collected in Kunming, Dali, Nujiang, Dehong, Yuxi, Lincang, Pu'er, Zhaotong, and Baoshan cities in Yunnan Province. Using Mycobacterium smegmatis mc²155 as the host bacterium, direct and enrichment methods were used to isolate Mycobacteriophages from the sample. The isolated Mycobacteriophages were subjected to biological characterization, whole genome sequencing, and genome analysis. Results: Three Mycobacteriophages were isolated from 186 samples, and one strain named YN1 was selected for the further study. The results showed that the optimal multiple number of YN1 infection was 0.1, and the growth incubation period was about 330 minutes. Environmental factors such as temperature, pH, UV and ethanol had certain effects on YN1.Genomic analysis showed that YN1 belongs to the linear double stranded DNA virus, while comparative genomics indicated that YN1 belongs to the mycobacterial Ⅰ cluster bacteriophage in taxonomy. Conclusion: This study isolated Mycobacteriophages from Yunnan Province for the first time. After identification and characterization, YN1 was identified as a new mycobacterial cluster Ⅰ bacteriophage, enriching the domestic mycobacterial bacteriophage library and providing a basis for further development and utilization of Mycobacteriophages.

Key words: Mycobacteriophages, Biology, Genome, Phenotype, Yunnan Province

中图分类号:

R378.91

杜毓, 张海鹏, 刘珂君, 石丽媛, 谭红丽, 王鹏. 分枝杆菌噬菌体在云南省的首次分离鉴定和特性研究[J]. 中国防痨杂志, 2025, 47(12): 1609-1615. doi: 10.19982/j.issn.1000-6621.20250248

Du Yu, Zhang Haipeng, Liu Kejun, Shi Liyuan, Tan Hongli, Wang Peng. First isolation, identification and characterization of Mycobacteriophages in Yunnan Province[J]. Chinese Journal of Antituberculosis, 2025, 47(12): 1609-1615. doi: 10.19982/j.issn.1000-6621.20250248

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感染复数	样本数(份)	效价[PFU/ml,M(Q₁, Q₃)]
0.001	3	6.00×10³(4.00×10³,10.0×10³)
0.01	3	6.00×10⁵(4.00×10⁵,8.00×10⁵)
0.1	3	1.20×10⁷(1.00×10⁷,1.40×10⁷)
1	3	4.00×10⁶(2.00×10⁶,4.00×10⁶)
10	3	1.60×10⁶(1.00×10⁶,1.80×10⁶)
100	3	2.40×10⁵(1.60×10⁵,2.40×10⁵)
1000	3	6.00×10⁴(4.00×10⁴,8.00×10⁴)

乙醇浓度(%)	样本数(份)	效价[PFU/ml,M(Q₁, Q₃)]	H值	P值
0	3	1.60×10⁷(1.50×10⁷,1.60×10⁷)	25.800	<0.001
10	3	8.00×10⁶(8.00×10⁶,10.00×10⁶)
25	3	8.00×10⁵(6.00×10⁵,8.00×10⁵)
50	3	0.00
75	3	0.00
95	3	0.00