大肠埃希菌裂解性噬菌体vB_EcoP-R1的特性及其抗细菌生物膜活性分析
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Characterization of Escherichia coli lytic phage vB_ EcoP-R1 and its anti-biofilm activity
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    摘要:

    摘要:目的从医 院污水中分离大肠埃希茵特异性噬菌体,分析其生物学特性,并检测其对细菌生物膜的裂解作用。方法. 通过双层琼脂平板法分离纯化噬菌体,透射电镜观察其形态,并测定其一步生长曲线、体外裂解曲线、温度及酸碱稳定性。全 基因测序分析噬菌体基因组组成及特性,结晶紫染色法评估噬菌体对宿主菌生物膜的清除效果。结果从医院污水中新分 离1株大肠埃希菌特异性噬菌体,命名为vB_ EoP-R1;透射电镜下显示形态与尾病毒目足病毒科噬菌体成员相似,在-20~ 50 C的温度范围内以及在pH值5.0~11.0酸碱条件下能稳定存活。vB_ EcoP-R1 基因组全长为40 875 bp,GC含量为49.94%, 不含毒力基因和耐药基因。vB_ _EcoP-R1 能够裂解临床分离的大肠埃希菌,且显示高度种属特异性。噬菌体作用于宿主菌预 成形生物膜4h后,对生物膜的清除率达到80.16%。结论vB_ EcoP-R1 是1株新的大肠埃希茵特异性噬菌体,裂解性强且种 属特异性高,可裂解宿主菌已形成的生物膜,有望用于大肠埃希菌感染的替代治疗。

    Abstract:

    Abstract: Objective To isolate Escherichia coli-specific bacteriophages from hospital waste water, analyze their biological characteristics and detect their cleavage effcts on bacterial biofilms.Methods The phages were isolated and purified by double-layer agar plate method and their morphology was observed by transmission electron microscopy. Their one-step growth curve, in vitro lysis curve, tem- perature and acid-base stability were determined. Whole genome sequencing was used to analyze the genome composition and characteristics of bacteriophages. The crystal violet staining was used to evaluate the scavenging effects of phage on host biofilms. Results A novel Escherichia coli-specific bacteriophage named vB_ EcoP-R1 was isolated from hospital sewage. Transmission electron microscopy revealed that the morphology of vB_ EcoP-R1 was similar to that of Podoviridae, a member of Caudovirales, and it could survive stably in the temperature range of -20 C to50 9C and pH 5.0 to 11.0. The total length of the vB_ EcoP-R1 genome was 40 875 bp and the GC content was 49.94%. There were no virulence genes and drug resistance genes in the phage genome. vB_ EcoP-R1 was capable of lysing clinically isolated Escherichia coli and showed high species and genus specificity. After the phage acted on the host bacteria preformned biofilm for 4 h, the clearance rate of the biofilm reached 80. 16%. Conclusion vB_ EcoP-R1 should be a novel Escherichia coli-specif-ic bacteriophage with strong lysis and high specificity for species and genus. The bacteriophage could lyse the biofilm formed by the host bacteria, and is expected to be used as an alternative treatment for Escherichia coli infection.

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石志新,李泽轩,张立涛,洪欣,刘径,赵哲,刘根焰.大肠埃希菌裂解性噬菌体vB_EcoP-R1的特性及其抗细菌生物膜活性分析[J].临床检验杂志,2024,42(4):261-266

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  • 收稿日期:2024-01-23
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  • 在线发布日期: 2024-06-14
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