■这项研究检查了导致对一线(利福平和异烟肼)抗性的遗传变化的模式和频率,氟喹诺酮类药物,和从Amhara地区精神圣水场所(HWS)的培养阳性肺结核(PTB)有症状的参与者中分离出的耐药结核分枝杆菌(MTB)的二线注射药物。
■从2019年6月到2020年3月,进行了横断面研究。对Amhara地区的HWS的PTB疑似参与者中的122株培养阳性MTB分离株进行了耐药性评估,以及赋予对利福平(RIF)抗性的特征基因突变,异烟肼(异烟肼),氟喹诺酮类药物(FLQs),以及使用GenoType®MTBDRplusVER2.0和GenoType®MTBDRslVER2.0的二线注射药物(SLID)。耐药MTB分离株按照制造商的方案进行Spoligotyped。
■导致RIF抗性的遗传变化(突变),INH,和FLQ在15/122(12.3%)中确定,20/122(16.4%),和5/20(25%)的MTB分离株,分别。在抗RIF中,rpoB/Ser531Lue(n=12,80%)最常见,其次是His526Tyr(6.7%)。在异烟肼抗性分离株中,katG/Ser315Thr1(n=19,95%)频率最高。在15例耐多药结核病中,大多数(n=12,80%)分离株在rpoB/Ser531Leu和katG/Ser315Thr1均有突变.用MTBDRslVER2.0测试了所有20种INH和/或RIF抗性分离株,产生5种具有rpoB/Ser531Lue基因突变的FLQs抗性分离株,katG/Ser315Thr1和gyrA/Asp94Ala基因。在20株Spoligotyped耐药MTB分离株中,大多数(n=11,55%)和6(30%)是SIT149/T3-ETH和SIT21/CAS1-Kili亚谱系,分别;它们是任何异烟肼抗性(单异型/多型)。在15株RIF耐药(RR/MDR-TB)分离株中,7是SIT149/T3-ETH,而6个是SIT21/CAS1-Kili亚谱系。在四个SIT21/CAS1-Kili谱系中检测到FLQ抗性。
■在当前的研究中,最常见的基因突变负责对异烟肼的抗性,RIF,并确定了FLQ。SIT149/T3-ETH和SIT21/CAS1-Kili构成了大多数耐药结核病(DR-TB)分离株。为了进一步了解基因变化/突变和相关基因型的完整谱,测序技术是必要的.
This study examined the patterns and frequency of genetic changes responsible for resistance to first-line (rifampicin and isoniazid), fluoroquinolones, and second-line injectable drugs in drug-resistant Mycobacterium tuberculosis (MTB) isolated from culture-positive pulmonary tuberculosis (PTB) symptomatic attendees of spiritual holy water sites (HWSs) in the Amhara region.
From June 2019 to March 2020, a cross-sectional study was carried out. A total of 122 culture-positive MTB isolates from PTB-suspected attendees of HWSs in the Amhara region were evaluated for their drug resistance profiles, and characterized gene mutations conferring resistance to rifampicin (RIF), isoniazid (INH), fluoroquinolones (FLQs), and second-line injectable drugs (SLIDs) using GenoType®MTBDRplus VER2.0 and GenoType®MTBDRsl VER2.0. Drug-resistant MTB isolates were Spoligotyped following the manufacturer\'s protocol.
Genetic changes (mutations) responsible for resistance to RIF, INH, and FLQs were identified in 15/122 (12.3%), 20/122 (16.4%), and 5/20 (25%) of MTB isolates, respectively. In RIF-resistant,
rpoB/Ser531Lue (n = 12, 80%) was most frequent followed by His526Tyr (6.7%). Amongst INH-resistant isolates, katG/Ser315Thr1 (n = 19, 95%) was the most frequent. Of 15 MDR-TB, the majority (n = 12, 80%) isolates had mutations at both
rpoB/Ser531Leu and katG/Ser315Thr1. All 20 INH and/or RIF-resistant isolates were tested with the MTBDRsl VER 2.0, yielding 5 FLQs-resistant isolates with gene mutations at
rpoB/Ser531Lue, katG/Ser315Thr1, and gyrA/Asp94Ala genes. Of 20 Spoligotyped drug-resistant MTB isolates, the majority (n = 11, 55%) and 6 (30%) were SIT149/T3-ETH and SIT21/CAS1-Kili sublineages, respectively; and they were any INH-resistant (mono-hetero/multi-). Of 15 RIF-resistant (RR/MDR-TB) isolates, 7 were SIT149/T3-ETH, while 6 were SIT21/CAS1-Kili sublineages. FLQ resistance was detected in four SIT21/CAS1-Kili lineages.
In the current study, the most common gene mutations responsible for resistance to INH, RIF, and FLQs were identified. SIT149/T3-ETH and SIT21/CAS1-Kili constitute the majority of drug-resistant TB (DR-TB) isolates. To further understand the complete spectrum of genetic changes/mutations and related genotypes, a sequencing technology is warranted.