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肯尼亚内罗毕初治结核病患者中结核分枝杆菌耐药突变的流行情况和检测。

Prevalence and detection of drug resistant mutations in Mycobacterium tuberculosis among drug naïve patients in Nairobi, Kenya.

机构信息

Microbiology Department, Kenyatta University, Nairobi, Kenya.

Kenya Medical Research Institute, Centre for Respiratory Disease and Research, Nairobi, Kenya.

出版信息

BMC Infect Dis. 2019 Mar 25;19(1):279. doi: 10.1186/s12879-019-3911-9.

Abstract

BACKGROUND

Tuberculosis (TB), an ancient scourge of humanity known for several thousands of years, is still a significant public health challenge in many countries today even though some progress has been made in recent years in controlling the disease. The study's aim was to determine the prevalence of mutations responsible for drug resistance in Mycobacterium tuberculosis among patients visiting selected health centers in Nairobi, Kenya.

METHODS

The cross-sectional study involved 132 TB positive patients visiting Mbagathi and Chandaria hospitals between September 2015 and August 2016. Sputum samples were collected from the participants and handled in a biosafety level 3 laboratory at the Kenya Medical Research Institute (KEMRI). Samples were decontaminated using N-Acetyl-L-Cysteine (NALC) - Sodium Hydroxide (NALC-NaOH), stained using Zeihl-Neelsen (ZN), and cultured in Mycobacterium Growth Indicator Tube (MGIT). DNA extracted from cultured isolates using Genolyse™ technique was subjected to Multiplex PCR amplification and reverse hybridization for detection of drug resistance mutations on rpoB, katG, inhA, gyrA, gyrB, rrs and eis genes using Hain Genotype MTBDRplus and MTBDRsl.

RESULTS

All 132 (100%) patients included in the study were culture positive for M. tuberculosis. Among them, 72 (54%) were male while the remaining 60 (46%) were female. The mean age of the patients was 26.4 ± 19.4 (SD) with a range of 18 to 60 years. Overall, the prevalence of the resistance to first and second-line TB drugs was 1.5% (2/132). Resistance to isoniazid (INH) was observed in 1 of 132 patients (0.8%), as was multi-drug resistant tuberculosis (MDR-TB), also at 0.8%. No resistance to fluoroquinolones (FQ) or kanamycin (KAN) was observed. The INH resistant strain had the katG mutations S315 T, while mutations detected for the MDR-TB were katG S513 T for INH, rpoB S531 L for rifampicin (RIF) and rrs G1484 T for cross-resistance to aminoglycosides/capreomycin (AG/CP).

CONCLUSIONS

Molecular analysis confirms transmission of the drug-resistant M. tuberculosis strains. The data suggested that there is homogeneity when it comes to the type of drug resistance and mutation that occurs in the region. This calls for intensified drug resistance surveillance and drug adherence among patients infected with TB.

摘要

背景

结核病(TB)是一种古老的人类疾病,已有数千年的历史,尽管近年来在控制该疾病方面取得了一些进展,但在许多国家仍然是一个重大的公共卫生挑战。本研究旨在确定肯尼亚内罗毕选定卫生中心就诊的结核病患者中导致结核分枝杆菌耐药的突变的流行率。

方法

这项横断面研究涉及 2015 年 9 月至 2016 年 8 月期间在 Mbagathi 和 Chandaria 医院就诊的 132 例结核分枝杆菌阳性患者。从参与者中采集痰液样本,并在肯尼亚医学研究所(KEMRI)的生物安全 3 级实验室中进行处理。使用 N-乙酰-L-半胱氨酸(NALC)-氢氧化钠(NALC-NaOH)对样本进行去污染,使用 Zeihl-Neelsen(ZN)进行染色,并在分枝杆菌生长指示剂管(MGIT)中进行培养。使用 Genolyse™技术从培养分离物中提取 DNA,然后使用 Hain Genotype MTBDRplus 和 MTBDRsl 对 rpoB、katG、inhA、gyrA、gyrB、rrs 和 eis 基因进行多重 PCR 扩增和反向杂交,以检测耐药性突变。

结果

所有纳入研究的 132 例(100%)患者均培养出结核分枝杆菌阳性。其中,72 例(54%)为男性,其余 60 例(46%)为女性。患者的平均年龄为 26.4±19.4(SD),年龄范围为 18 至 60 岁。总体而言,一线和二线抗结核药物耐药率为 1.5%(2/132)。132 例患者中有 1 例(0.8%)对异烟肼(INH)耐药,1 例(0.8%)对耐多药结核病(MDR-TB)耐药。未观察到氟喹诺酮类(FQ)或卡那霉素(KAN)耐药。耐 INH 株存在 katG S315T 突变,而 MDR-TB 检测到的突变分别为 INH 的 katG S513T、利福平(RIF)的 rpoB S531L 和交叉耐药性氨基糖苷类/卷曲霉素(AG/CP)的 rrs G1484T。

结论

分子分析证实了耐药结核分枝杆菌株的传播。数据表明,该地区发生的耐药类型和突变具有同质性。这需要加强耐药性监测和患者对结核病的药物依从性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f697/6434623/a712c203507b/12879_2019_3911_Fig1_HTML.jpg

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