类鼻疽伯克霍尔德菌标准菌株的建立与评价

刘旭玲; 方艳梅; 京舒娉; 董莉; 翁俊嘉; 吴佳韩; 崔莹; 林毅雄; 黄辉涛; 黄文燕

doi:10.3784/jbjc.202510200702

类鼻疽伯克霍尔德菌标准菌株的建立与评价

Establishment and evaluation of standard strains of Burkholderia pseudomallei

摘要

摘要:
目的依据《病原微生物菌（毒）种国家标准株评价技术标准》（WS/T 812—2022），构建类鼻疽伯克霍尔德菌标准菌株。
方法以ZH20BC01为研究对象，通过菌株鉴定、形态特征分析、动力学试验、生化特征分析、抗菌药物敏感性对其菌株进行系统的病原学研究。多次传代后，观察和比较不同代次间细菌生化、药敏特性以及分子遗传特征，对ZH20BC01进行稳定性评价，明确其病原微生物学特征。
结果菌落形态、革兰染色、电镜实验、质谱鉴定、16S rDNA测序、生化鉴定等结果证明ZH20BC01菌株具备类鼻疽伯克霍德尔菌的典型特征。除对阿米卡星、多粘菌素B具有耐药性，ZH20BC01对多数受试抗生素具有敏感性表型。ZH20BC01的G1、G5、G10三代菌的生化特征和药敏特征无明显差别，分子遗传特征稳定。
结论初步建立并评价了类鼻疽伯克霍尔德菌的标准菌株，为类鼻疽病的疾病控制、临床诊断、教学科研、疫苗研发提供菌株资源。

Abstract:
Objective According to the Technical Requirements for Evaluation of National Standard Strains of Pathogenic Microorganisms(WS/T 812—2022), construct a standard strain of Burkholderia pseudomallei.
Methods A systematic pathogenic study was conducted on ZH20BC01 through strain identification, morphological feature, motility characteristics, biochemical characteristics, antibiotic susceptibility. The passage stability of the ZH20BC01 were further determined by comparing and analyzing the differences in biochemical characteristics,susceptibility to antibiotics and genetic stability between G1, G5 and G10 strains.
Results The results of colony morphology, Gram staining, Transmission Electron Microscope studies, Matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) identification, 16S rDNA sequencing and biochemical testing demonstrate that the ZH20BC01 strain has typical characteristics of Burkholderia pseudomallei. Except for antibiotics such as amikacin and polymyxin B, ZH20BC01 showed a susceptible phenotype to most of the tested antibiotics. There is no significant difference in biochemical characteristics, antibiotic susceptibility and molecular genetic characteristics among the G1, G5, and G10 generations of ZH20BC01 bacteria, indicating good passage stability.
Conclusion A standard strain of Burkholderia pseudomallei was established and evaluated, which could be used in disease control,clinical diagnosis, teaching and scientific research and vaccine development in China.

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