Etiological identification and biological characteristics of recurrent melioidosis: report of 1 case

Wenbo YANG; Na'an ZHANG; Chenglong RAO; Yupei XIA; Meijuan ZHANG; Xiao LI; Jingmin YAN; Hai CHEN; Qian LI; Xuhu MAO

doi:10.16016/j.1000-5404.202105139

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Etiological identification and biological characteristics of recurrent melioidosis: report of 1 case

Basic Medicine | 更新时间：2026-01-09

- Etiological identification and biological characteristics of recurrent melioidosis: report of 1 case
- Journal of Third Military Medical University Vol. 43, Issue 22, Pages: 2441-2448(2021)
- 作者机构：
  
  1.400038    重庆，陆军军医大学（第三军医大学）药学与检验医学系临床微生物与免疫学教研室
  2.430070   湖北，华中农业大学生命科学技术学院
  3.572000   海南，三亚市人民医院检验科
- 作者简介：
- 基金信息：
  
  the General Program of National Natural Science Foundation of China(81971907)
- DOI：10.16016/j.1000-5404.202105139
  CLC： R372;R378.99;R515.9
- Received：26 May 2021，
  
  Revised：2021-08-31，
  
  Published：30 November 2021
- 稿件说明：
移动端阅览
Wenbo YANG, Na'an ZHANG, Chenglong RAO, et al. Etiological identification and biological characteristics of recurrent melioidosis: report of 1 case[J]. Journal of Third Military Medical University, 2021, 43(22): 2441-2448.
DOI：

Wenbo YANG, Na'an ZHANG, Chenglong RAO, et al. Etiological identification and biological characteristics of recurrent melioidosis: report of 1 case[J]. Journal of Third Military Medical University, 2021, 43(22): 2441-2448. DOI： 10.16016/j.1000-5404.202105139.

摘要

目的

通过对1例复发类鼻疽进行系统病原学鉴定及生物学特性研究，分析和探讨类鼻疽菌长期滞留机体可能的适应宿主机制。

方法

对临床收集的来源于同一个患者时隔5年分离到的两株类鼻疽菌，进行微生物学鉴定、全基因组测序，并使用相关软件对测序数据进行多位点序列分型（multilocus sequence typing，MLST）、同源比对进化树、单核苷酸多态性（single nucleotide polymorphism，SNP）以及插入缺失（insertion-deletion，Indel）分析；进一步在感染肺上皮A549细胞模型中比较感染不同时间细胞内细菌载量变化；双向电泳-质谱技术分析两株细菌在蛋白水平的表达变化。

结果

分离到的两株类鼻疽菌，全基因组均约7.12 Mb，COG功能注释编码蛋白大致约5 000个；MLST分型均为ST58型；同源比对进化树分析显示它们位于同一分支且与其他分离自中国的类鼻疽菌亲缘关系较近；SNP/Indel分析显示复发株有114个SNP位点、55个Indel位点；与原发株相比，双向电泳-质谱结果表明复发株共有68个蛋白的表达水平发生了显著的变化；同时复发株感染A549后细胞内细菌载量明显增加（

0.05）。

结论

以上复发菌株的基因组学、蛋白组学以及胞内感染增殖能力的系统研究提示类鼻疽菌可能在机体长期的选择压力下微进化，通过对细菌基因转录表达、分泌系统、耐药以及能量代谢等方面的调控更加适应其在宿主体内的生存而实现长期滞留。

Abstract

Objective

To analyze and explore the possible adaptive mechanisms that Burkholderia

pseudomallei

(B.

pseudomallei

) could long-term reside in the host cells based on the systematic etiological identification and biological characteristics of a patient with recurrent melioidosis.

Methods

The 2 B.

pseudomallei

strains isolated from the same patient at an interval of 5 years were performed for microbiological identification and genome sequencing

and the sequencing data were further analyzed by multilocus sequence typing (MLST)

phylogenetic tree analysis of homologous alignment

single nucleotide polymorphism (SNP) analysis and insertion-deletion (Indel) analysis

respectively. Furthermore

the intracellular survival of the 2 clinical isolates of B.

pseudomallei

were examined at different times by counting the viable bacteria in the A549 cells. Two-dimensional electrophoresis (2-DE) and mass spectrometry was employed to analyze the protein expression of 2 strains of bacteria.

Results

The 2 clinical isolates had a whole genome of about 7.12 Mb

and up about 5 000 Clusters of Orthologous Groups (COG) annotations

and was identified as sequence type (ST) 58 by MLST analysis. Phylogenetic analysis of homologous alignment showed that they were located in the same branch and were closely related to other B.

pseudomallei

isolates from China. SNP/Indel analysis indicated that there were 114 SNP sites and 55 Indel sites in the recurrent strain. The results of 2-DE displayed that the expression levels of 68 proteins in the recurrent strain were significantly changed compared with the primary strain. The intracellular bact

erial load of recurrent strain was significantly increased compared to the primary strain in A549 cells(

0.05).

Conclusion

Based on a series of studies with genomics

proteomics and ability of intracellular survival of the 2 clinical isolates of

pseudomallei

the bacterium may evolve slightly under the long-term selective pressure of the organism

and adapt to the host environment for its own survival through the regulation of bacterial transcription

secretion system

drug resistance and energy metabolism.

关键词

Keywords

references

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