Department of Pulmonology and Respiratory Medicine; Laboratory of Tuberculosis, Institute of Tropical Disease, Airlangga University, Surabaya, Indonesia.
Department of Pulmonology and Respiratory Medicine, Airlangga University, Surabaya, Indonesia.
Int J Mycobacteriol. 2020 Oct-Dec;9(4):380-390. doi: 10.4103/ijmy.ijmy_175_20.
The disease severity in pulmonary Multidrug-resistant tuberculosis (MDR-TB) varies from mild to severe, which is determined by host and pathogen virulence factors. The difference of symptoms felt by TB patients were interesting to investigate in discovering whether its the human immune response or bacteria's virulence gene that plays the role. The aim of this research was to analyze association between disease severity degree of pulmonary MDR-TB patients with Single nucleotide polymorphisms (SNPs) found in toll-like receptors (TLRs) gene.
Blood samples were obtained from pulmonary MDR-TB patients in Dr. Soetomo Hospital, Surabaya, Indonesia. Polymerase chain reaction (PCR) multiplex and target SNPs were analyze using DigiTag2 assay. The variant of esxA gene was determined using PCR and sequencing. Severity degree was determined by chest X-ray, the lesions were scored according to their severity, score of =2.5 ranking as mild, 2.5-6 as moderate and =6 as severe. Association level between SNP in TLRs gene degree of pulmonary MDR-TB was analyzed using Chi-square test. Bonferroni correction for multiple comparison was used to anticipate genotyping error.
A total of 22 MDR-TB patients were classified into severe degree group, while 16 patients were moderate/mild degree. SNPs in encoding gene of TLRs were mostly found in intron, specifically in TLR-1, TLR-2, and TLR-6. HWE P value in rs5743572 was 0.841; in rs3804100 was 0.0176; and in rs5743808 was 0.562. Association analysis between SNP in TLRs genes and degree of disease revealed significant association in rs5743572, SNP of TLR-1, with P < 0.05; odds ratio [OR] = 11.67 (95% confidence interval [CI]: 3.94-34.52); rs3804100, SNP of TLR-2 had P < 0.05; OR = 37.59 (95% CI: 9.30-151.88); and rs5743808, the SNP of TLR-6 had P < 0.05; OR = 31.5 (95% CI: 8.60-115.34).
We concluded that SNPs in TLR-1, TLR-2, and TLR-6 of pulmonary MDR-TB patients was found to have an association with disease severity. TLRs polymorphism had significant association was present in TLR-1 rs5743572 in intron, TLR-2 rs3804100 in exon, and TLR-6 rs5743808 in exon and among MDR-TB isolates from patients with pulmonary MDR-TB of severe and moderate/mild degree.
肺部耐多药结核病（MDR-TB）的疾病严重程度从轻到重不等，这取决于宿主和病原体的毒力因素。研究结核病患者的症状差异很有趣，这有助于发现是人类免疫反应还是细菌的毒力基因在起作用。本研究旨在分析肺部 MDR-TB 患者疾病严重程度与 Toll 样受体（TLRs）基因中发现的单核苷酸多态性（SNPs）之间的关联。
从印度尼西亚泗水苏加诺哈达医院的肺部 MDR-TB 患者中获得血液样本。使用聚合酶链反应（PCR）多重和靶向 SNPs 分析 DigiTag2 测定法。使用 PCR 和测序确定 esxA 基因的变体。通过胸部 X 射线确定严重程度，根据严重程度对病变进行评分，=2.5 为轻度，2.5-6 为中度，>=6 为重度。使用卡方检验分析 TLRs 基因中 SNP 与肺部 MDR-TB 严重程度的关联水平。使用 Bonferroni 校正进行多重比较以预测基因分型错误。
总共 22 例 MDR-TB 患者被分为严重程度组，而 16 例患者为中度/轻度。TLRs 基因编码区的 SNPs 主要存在于内含子中，特别是 TLR-1、TLR-2 和 TLR-6。rs5743572 的 HWE P 值为 0.841；rs3804100 为 0.0176；rs5743808 为 0.562。TLRs 基因中 SNP 与疾病程度的关联分析显示，rs5743572（TLR-1 的 SNP）与 P<0.05 之间存在显著关联；比值比[OR] = 11.67（95%置信区间[CI]：3.94-34.52）；rs3804100（TLR-2 的 SNP）与 P<0.05；OR = 37.59（95% CI：9.30-151.88）；rs5743808（TLR-6 的 SNP）与 P<0.05；OR = 31.5（95% CI：8.60-115.34）。
我们得出结论，肺部 MDR-TB 患者 TLR-1、TLR-2 和 TLR-6 中的 SNPs 与疾病严重程度相关。TLRs 多态性与 TLR-1 rs5743572 在内含子、TLR-2 rs3804100 在外显子和 TLR-6 rs5743808 在外显子中的显著关联有关，并且与严重和中度/轻度肺部 MDR-TB 患者的 MDR-TB 分离株有关。
