背景:毒素-抗毒素系统是一种遗传元件,在结核分枝杆菌(MTB)中高度存在,结核病的病原体。毒素-抗毒素系统包括毒素蛋白和抗毒素蛋白或非编码RNA,它们彼此相互作用并抑制毒素活性。结核分枝杆菌比非结核杆菌和其他微生物具有更多种类的TA基因座,包括VapBC,希格巴,MazEF,ParDE,RelBE,MbcTA,PemIK,DarTG,门塔,一个三方II型TAC陪伴系统,和假设的蛋白质。
目的:该研究旨在证明临床样本中结核分枝杆菌菌株中编码毒素-抗毒素系统的基因。
方法:收集肺结核和肺外肺结核临床标本,和涂片显微镜(Ziehl-Neelsen染色)进行检测高杆菌(3+)计数,然后进行核酸扩增测定。细菌培养和生长测定,基因组DNA提取,并进行了聚合酶链反应。
结果:在显微镜涂片中通过3确定了阳性PTB和EPTB样品[20],NAAT法测定痰中结核杆菌总数为8.0×1005,组织脓肿为1.3×1004CFU/ml。此外,从培养物中提取基因组DNA,以及Rv1044和Rv1045基因在624和412个碱基对中的扩增(在梯级中在600-700和400-500之间),分别,在H37Rv和临床样本中观察到。
结论:已经发现Rv1044和Rv1045是具有624和882个碱基对的假设蛋白质,属于AbiEi/AbiEii家族的毒素-抗毒素基因座。此外,TA编码基因座基因的显著鉴定可能有助于研究耐多药和广泛耐药结核病.
BACKGROUND: The toxin-antitoxin system is a genetic element that is highly present in Mycobacterium tuberculosis (MTB), the causative agent of tuberculosis. The toxin-antitoxin system comprises toxin protein and antitoxin protein or non-encoded RNA interacting with each other and inhibiting toxin activity. M. Tuberculosis has more classes of TA loci than non-tubercle bacilli and other microbes, including VapBC, HigBA, MazEF, ParDE, RelBE, MbcTA, PemIK, DarTG, MenTA, one tripartite type II TAC chaperone system, and hypothetical proteins.
OBJECTIVE: The study aims to demonstrate the genes encoded toxin-antitoxin system in mycobacterium tuberculosis strains from clinical samples.
METHODS: The pulmonary and extra-pulmonary tuberculosis clinical samples were collected, and smear microscopy (Ziehl-Neelsen staining) was performed for the detection of high bacilli (3+) count, followed by nucleic acid amplification assay. Bacterial culture and growth assay, genomic DNA extraction, and polymerase chain reaction were also carried out.
RESULTS: The positive PTB and EPTB samples were determined by 3+ in microscopy smear and the total count of tubercle bacilli determined by NAAT assay was 8.0×1005 in sputum and 1.3×1004 CFU/ml in tissue abscess. Moreover, the genomic DNA was extracted from culture, and the amplification of Rv1044 and Rv1045 genes in 624 and 412 base pairs (between 600-700 and 400-500 in ladder), respectively, in the H37Rv and clinical samples was observed.
CONCLUSIONS: It has been found that Rv1044 and Rv1045 are hypothetical proteins with 624 and 882 base pairs belonging to the AbiEi/AbiEii family of toxin-antitoxin loci. Moreover, the significant identification of TA-encoded loci genes may allow for the investigation of multidrugresistant and extensively drug-resistant tuberculosis.