Hadizadeh Tasbiti Alireza, Yari Shamsi, Siadat Seyed Davar, Karimipoor Morteza, Badmasti Farzad, Masoumi Morteza, Abdolrahimi Farid, Khanipour Sharareh, Hassanzadeh Seyed Mehdi, Ghalami Nobar Mostafa, Yari Fatemeh
Tuberculosis and Pulmonary Research Dept. Pasteur Institute of Iran, Tehran, Iran.
Microbiology Research Center (MRC), Pasteur Institute of Iran, Tehran, Iran.
Biotechnol Rep (Amst). 2021 May 29;30:e00641. doi: 10.1016/j.btre.2021.e00641. eCollection 2021 Jun.
Tuberculosis (TB) is a sizable public health threat in the world. This study was conducted to determine the differential protein composition between susceptible and MDRTB strains. Tuberculosis proteins were extracted by Triton™ X-114 and ammonium sulfate. Two-dimensional gel electrophoresis protein spots were selected for identification by mass spectrometry and mRNA expression levels were measured by real- time PCR. 2DE-Western blot and T cell epitope prediction for identified proteins were made by the IEDB server. The result shows at least six protein spots (Rv0147, Rv3597c, Rv0379, Rv3699, Rv1392 and Rv0443) were differentially expressed in MDRTB isolates. However, difference in mRNA gene expression was not found in the six mRNA genes. 2DE-Western blot procedures indicated strong reaction against MDRTB proteins corresponds to 13, 16 and 55 kDa areas that might be used as new diagnostic tools. In conclusion, these MDRTB proteins identified in this study could be reliable TB diagnostic candidates or therapeutic targets.
结核病（TB）是全球范围内一项重大的公共卫生威胁。本研究旨在确定敏感菌株与耐多药结核菌株之间的差异蛋白质组成。采用Triton™ X - 114和硫酸铵提取结核蛋白。通过质谱对二维凝胶电泳蛋白点进行选择鉴定，并通过实时PCR测定mRNA表达水平。利用IEDB服务器对鉴定出的蛋白质进行二维电泳 - 蛋白质印迹分析和T细胞表位预测。结果显示，至少有六个蛋白点（Rv0147、Rv3597c、Rv0379、Rv3699、Rv1392和Rv0443）在耐多药结核分枝杆菌分离株中差异表达。然而，在这六个mRNA基因中未发现mRNA基因表达的差异。二维电泳 - 蛋白质印迹分析程序表明，针对耐多药结核分枝杆菌蛋白的强烈反应对应于13、16和55 kDa区域，这些区域可能用作新的诊断工具。总之，本研究中鉴定出的这些耐多药结核分枝杆菌蛋白可能是可靠的结核病诊断候选物或治疗靶点。
