高通量测序筛选结核分枝杆菌耐药株差异基因  

Screening for differentially expressed genes of Mycobacterium tuberculosis drug resistant strains by high-throughput sequencing

作　　者：李劭源[1,2] 张国良[3] 文玉欣 毛冬婷 曹开源[1,2] 陈心春[3] 曾谷城[1,2] 

机构地区：[1]中山大学中山医学院微生物学教研室,广东广州510080 [2]中山大学热带病防治研究教育部重点实验室,广东广州510080 [3]广东医学院附属深圳市第三人民医院肝病研究所,广东深圳518112 [4]暨南大学第二临床医学院深圳市人民医院胸外科,广东深圳518020

出　　处：《热带医学杂志》2014年第5期547-550,共4页Journal of Tropical Medicine

基　　金：国家自然科学基金(C080102);国家自然科学基金委员会与美国国立卫生研究院合作研究项目(812111408)

摘　　要：目的比较结核分枝杆菌中敏感株、多耐药株、广泛耐药株三者间基因表达的差异,为进一步针对耐药菌株的研究提供方向。方法从深圳市第三人民医院收集敏感株、多耐药株、广泛耐药株各1株,提取菌株RNA,然后构建cDNA文库,使用Genome Analyzer IIx高通量测序仪进行全基因组测序,经过筛选比对后得到测序结果 ,再比较3个菌株间的基因表达差异。结果高通量测序后共得到基因3 968个。将耐药株与敏感株的基因进行比较,选取表达差异高于10倍的基因,筛选得到多耐药株差异基因16个、广泛耐药株差异基因10个。这些表达差异基因大部分与毒素-抗毒素系统、PE/PPE家族蛋白、金属转运蛋白相关。还在多耐药株中发现了一系列高表达的前噬菌体基因及噬菌体整合相关的基因。结论通过高通量测序筛选得到一系列耐药株差异表达基因,为未来对耐药菌株的研究提供依据。Objective We aimed to compare the differences of gene expression levels among sensitive strain （S）, multi-drug resistant strain （MDR） and extensive-drug resistant strain （XDR） and provide direction for further research of drug resistant strains. Methods S, MDR and XDR strains were collected from the Third People＇s Hospital of Shenzhen. RNA samples of these strains were used to construct cDNA libraries. Then the cDNA libraries were sequenced by Genome Analyzer IIx to obtain the whole transcriptome sequences. The gene expression differences among 3 strains were compared. Results 3 968 genes were obtained after High-throughput sequencing. Gene expression levels were compared between drug resistant strains and sensitive strain. Genes which were 10 folds higher in expression levels in drug resistant strains were chosen. 16 differentially expressed genes were identified in MDR and 10 in XDR. Most of these genes were associated with toxin-antitoxin system, PE/PPE family protein and metal transporters. A series of PhiRvl phage and integrating genes were also found. Conclusion We obtained a series of differentially expressed genes related to drug resistance of tuberculosis, providing proofs for future research of drug resistance of Mycobacterium tuberculosis.

关 键 词：结核分枝杆菌 耐药株 高通量测序 

分 类 号：R378.911[医药卫生—病原生物学]