Wang Zhirui, Sun Rui, Mu Cheng, Wang Chunhua, Zhao Hui, Jiang Lina, Ju Hanfang, Dai Wenxi, Zhang Fan
Tuberculosis Reference Laboratory, Tianjin Center for Tuberculosis Control, Tianjin, People's Republic of China.
Infect Drug Resist. 2022 Apr 13;15:1793-1803. doi: 10.2147/IDR.S361635. eCollection 2022.
Tuberculosis (TB) caused by remains a global concern. This study aimed to determine the molecular characteristics of fluoroquinolone-resistant and multidrug-resistant strains using whole-genome sequencing to predict drug resistance in in Tianjin, China, which has not been established previously.
Twenty-one fluoroquinolone-resistant and multidrug-resistant strains were isolated from sputum samples. Phenotypic drug resistance against 12 anti-tuberculosis drugs was determined using drug susceptibility testing. Whole-genome sequencing was performed to predict drug resistance in based on genome regions associated with drug resistance. The sensitivity of whole-genome sequencing for predicting drug resistance was calculated based on phenotypic drug susceptibility testing information.
Among the 21 isolates, mutations in 15 genome regions associated with drug resistance, including for rifampicin; and promoter for isoniazid; and for ofloxacin and moxifloxacin; for streptomycin; for streptomycin, amikacin, kanamycin and capreomycin; and for pyrazinamide; , and for ethambutol; for protionamide; and for para-aminosalicylic acid, were detected. Compared with traditional drug susceptibility testing results, the sensitivities for whole-genome sequencing of rifampin, isoniazid, ofloxacin, moxifloxacin, streptomycin, ethambutol, pyrazinamide, kanamycin, and amikacin resistance were 100%, 90.48%, 95.24%, 92.86%, 95.27%, 85.71%, 66.67%, 50%, and 50%, respectively. The sensitivities for whole-genome sequencing of capreomycin, protionamide, and para-aminosalicylic acid were not calculated because only one isolate showed phenotypic drug resistance. Mutations determined in drug susceptibility-associated genes can explain phenotypic drug resistance in most isolates. Notably, these mutations were absent in certain drug-resistant isolates, indicating other drug resistance mechanisms.
Whole-genome sequencing represents an effective diagnostic tool for fluoroquinolone-resistant and multidrug-resistant TB though it has some obstacles. Whole-genome sequencing should be used to predict drug resistance prior to performing traditional phenotypic drug susceptibility testing in Tianjin, China.
由[具体病原体]引起的结核病仍是全球关注的问题。本研究旨在利用全基因组测序确定耐氟喹诺酮和耐多药[病原体名称]菌株的分子特征，以预测中国天津[病原体名称]的耐药性，此前尚未开展过此类研究。
从痰液样本中分离出21株耐氟喹诺酮和耐多药[病原体名称]菌株。使用药敏试验确定对12种抗结核药物的表型耐药性。基于与耐药性相关的基因组区域进行全基因组测序，以预测[病原体名称]的耐药性。根据表型药敏试验信息计算全基因组测序预测耐药性的敏感性。
在21株分离株中，检测到15个与耐药性相关的基因组区域发生突变，包括利福平的[相关基因]；异烟肼的[相关基因]和启动子；氧氟沙星和莫西沙星的[相关基因]；链霉素的[相关基因]；链霉素、阿米卡星、卡那霉素和卷曲霉素的[相关基因]；吡嗪酰胺的[相关基因]；乙胺丁醇的[相关基因]、[相关基因]和[相关基因]；丙硫异烟胺的[相关基因]；对氨基水杨酸的[相关基因]。与传统药敏试验结果相比，全基因组测序对利福平、异烟肼、氧氟沙星、莫西沙星、链霉素、乙胺丁醇、吡嗪酰胺、卡那霉素和阿米卡星耐药性的敏感性分别为100%、90.48%、95.24%、92.86%、95.27%、85.71%、66.67%、50%和50%。由于只有1株分离株表现出表型耐药性，因此未计算全基因组测序对卷曲霉素、丙硫异烟胺和对氨基水杨酸的敏感性。在药敏相关基因中确定的突变可以解释大多数分离株的表型耐药性。值得注意的是，某些耐药分离株中不存在这些突变，表明存在其他耐药机制。
尽管全基因组测序存在一些障碍，但它是耐氟喹诺酮和耐多药结核病的一种有效诊断工具。在中国天津，应在进行传统表型药敏试验之前，使用全基因组测序来预测耐药性。
