National Tuberculosis Reference Laboratory, TB/HIV Directorate, Ethiopian Public Health Institute, Addis Ababa, Ethiopia.
Tropical and Infectious Diseases, Aklilu Lemma Institute of Pathobiology, Addis Ababa University, Addis Ababa, Ethiopia.
PLoS One. 2023 Jul 28;18(7):e0284363. doi: 10.1371/journal.pone.0284363. eCollection 2023.
Worldwide, tuberculosis (TB) affects about one million children every year. The burden of the disease is higher in developing countries. However, there is limited information on the lineages and drug sensitivity patterns of Mycobacterium tuberculosis (M. tuberculosis) infecting children in these countries, including Ethiopia. Thus, this study aimed to characterize the different lineages of the M. tuberculosis complex causing childhood pulmonary tuberculosis and evaluate the drug-sensitivity patterns to the first-line anti-TB drugs.
A total of 54 stored cultures were used in this study. The region of difference 9 (RD9) based polymerase chain reaction (PCR) and spoligotyping were employed for the identification of the isolates at the species and lineages level respectively. Lineage identification was done by using the pre-existing database. Identification of clustering of the spoligotype patterns was by using the SPOLIDB3-based model. The result was retrieved by the most probable family format. Furthermore, the phenotypic, and genotypic drug-sensitivity test (DST) was performed using Mycobacterium Growth Indicator Tube (MGIT™ 960) and GenoTypeMTBDRplus assay respectively. Data analysis was done using SPSS version 27 software.
Spoligotyping produced 39 interpretable results for M. tuberculosis. The majority (74.4%) of them were clustered into 7 groups, while the rest (25.6%) were single. The Euro-American (EA) lineage was the predominant lineage (64.1%) followed by the East-African Indian (EAI) (30.8%) and M. Africanum (5.1%) lineages. The most predominant subtypes were SIT37 (15.4%), SIT149 (12.8%), SIT25 (7.7%), and SIT53 (7.7%). Furthermore, of the identified SITs, T1 and CAS families consisted of 38.5% and 28.2% of the lineages respectively. Drug susceptibility was 91.9% by phenotypic method and 97.4% by molecular assay. The overall prevalence of any resistance was 7.8% and there was a single MDR-TB.
Many of the isolates belong to the modern lineages (Euro American) representing the most common circulating strains in the country. More importantly, despites the tiny isolates tested, drug resistance is low. To fully describe the molecular epidemiology of MTBC lineages in children, we recommend a prospective large-scale study.
全世界每年约有 100 万儿童感染结核病(TB)。发展中国家的疾病负担更高。然而,关于这些国家儿童感染结核分枝杆菌(M. tuberculosis)的谱系和药物敏感性模式的信息有限,包括埃塞俄比亚。因此,本研究旨在描述引起儿童肺结核的结核分枝杆菌复合体的不同谱系,并评估一线抗结核药物的药物敏感性模式。
本研究共使用了 54 株储存培养物。基于差异区 9(RD9)的聚合酶链反应(PCR)和 spoligotyping 分别用于鉴定分离株的种和谱系水平。通过现有的数据库进行谱系鉴定。 spoligotype 模式聚类的鉴定通过基于 SPOLIDB3 的模型进行。结果以最可能的家族格式检索。此外,使用分枝杆菌生长指示管(MGIT™960)和 GenoTypeMTBDRplus 检测分别进行表型和基因型药物敏感性试验(DST)。数据分析使用 SPSS 版本 27 软件进行。
spoligotyping 产生了 39 个可解释的结核分枝杆菌结果。其中大多数(74.4%)聚类为 7 组,其余(25.6%)为单一组。欧洲-美洲(EA)谱系是最主要的谱系(64.1%),其次是东非-印度(EAI)(30.8%)和 M. Africanum(5.1%)谱系。最主要的亚型是 SIT37(15.4%)、SIT149(12.8%)、SIT25(7.7%)和 SIT53(7.7%)。此外,在所鉴定的 SIT 中,T1 和 CAS 家族分别包含 38.5%和 28.2%的谱系。表型方法的药物敏感性为 91.9%,分子检测法为 97.4%。总耐药率为 7.8%,且存在单一致病菌。
许多分离株属于现代谱系(欧洲-美洲),代表了该国最常见的循环菌株。更重要的是,尽管测试的分离株很小,但耐药率很低。为了全面描述儿童结核分枝杆菌复合体的分子流行病学,我们建议进行一项前瞻性的大规模研究。