Xie Yingda L, Chakravorty Soumitesh, Armstrong Derek T, Hall Sandra L, Via Laura E, Song Taeksun, Yuan Xing, Mo Xiaoying, Zhu Hong, Xu Peng, Gao Qian, Lee Myungsun, Lee Jongseok, Smith Laura E, Chen Ray Y, Joh Joon Sung, Cho YoungSoo, Liu Xin, Ruan Xianglin, Liang Lili, Dharan Nila, Cho Sang-Nae, Barry Clifton E, Ellner Jerrold J, Dorman Susan E, Alland David
From the Tuberculosis Research Section, Laboratory of Clinical Infectious Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda (Y.L.X., L.E.V., R.Y.C., C.E.B.), and Johns Hopkins University School of Medicine, Baltimore (D.T.A., S.E.D.) - both in Maryland; the Center for Emerging and Re-Emerging Pathogens, Rutgers New Jersey Medical School, Newark (S.C., L.E.S., N.D., D.A.); Boston Medical Center and Boston University School of Medicine, Boston (S.L.H., J.J.E.); the International Tuberculosis Research Center, Changwon (T.S., M.L., J.L., S.-N.C.), and the National Medical Center (J.S.J.), Seoul Metropolitan Seobuk Hospital (Y.C.), and the Department of Microbiology, College of Medicine, Yonsei University (S.-N.C.), Seoul - all in South Korea; Henan Provincial Chest Hospital (X.Y., X.M., X.L., X.R., L.L.) and Sino-U.S. Tuberculosis Research Collaboration (H.Z.), Zhengzhou, and Key Laboratory of Medical Molecular Virology of Ministries of Education and Health, School of Basic Medical Science, Fudan University, Shanghai (P.X., Q.G.) - all in China; and the Institute of Infectious Disease and Molecular Medicine and Department of Pathology, Faculty of Health Sciences, University of Cape Town, Cape Town, South Africa (C.E.B.).
N Engl J Med. 2017 Sep 14;377(11):1043-1054. doi: 10.1056/NEJMoa1614915.
Fluoroquinolones and second-line injectable drugs are the backbone of treatment regimens for multidrug-resistant tuberculosis, and resistance to these drugs defines extensively drug-resistant tuberculosis. We assessed the accuracy of an automated, cartridge-based molecular assay for the detection, directly from sputum specimens, of Mycobacterium tuberculosis with resistance to fluoroquinolones, aminoglycosides, and isoniazid.
We conducted a prospective diagnostic accuracy study to compare the investigational assay against phenotypic drug-susceptibility testing and DNA sequencing among adults in China and South Korea who had symptoms of tuberculosis. The Xpert MTB/RIF assay and sputum culture were performed. M. tuberculosis isolates underwent phenotypic drug-susceptibility testing and DNA sequencing of the genes katG, gyrA, gyrB, and rrs and of the eis and inhA promoter regions.
Among the 308 participants who were culture-positive for M. tuberculosis, when phenotypic drug-susceptibility testing was used as the reference standard, the sensitivities of the investigational assay for detecting resistance were 83.3% for isoniazid (95% confidence interval [CI], 77.1 to 88.5), 88.4% for ofloxacin (95% CI, 80.2 to 94.1), 87.6% for moxifloxacin at a critical concentration of 0.5 μg per milliliter (95% CI, 79.0 to 93.7), 96.2% for moxifloxacin at a critical concentration of 2.0 μg per milliliter (95% CI, 87.0 to 99.5), 71.4% for kanamycin (95% CI, 56.7 to 83.4), and 70.7% for amikacin (95% CI, 54.5 to 83.9). The specificity of the assay for the detection of phenotypic resistance was 94.3% or greater for all drugs except moxifloxacin at a critical concentration of 2.0 μg per milliliter (specificity, 84.0% [95% CI, 78.9 to 88.3]). When DNA sequencing was used as the reference standard, the sensitivities of the investigational assay for detecting mutations associated with resistance were 98.1% for isoniazid (95% CI, 94.4 to 99.6), 95.8% for fluoroquinolones (95% CI, 89.6 to 98.8), 92.7% for kanamycin (95% CI, 80.1 to 98.5), and 96.8% for amikacin (95% CI, 83.3 to 99.9), and the specificity for all drugs was 99.6% (95% CI, 97.9 to 100) or greater.
This investigational assay accurately detected M. tuberculosis mutations associated with resistance to isoniazid, fluoroquinolones, and aminoglycosides and holds promise as a rapid point-of-care test to guide therapeutic decisions for patients with tuberculosis. (Funded by the National Institute of Allergy and Infectious Diseases, National Institutes of Health, and the Ministry of Science and Technology of China; ClinicalTrials.gov number, NCT02251327 .).
氟喹诺酮类药物和二线注射用药物是耐多药结核病治疗方案的核心,对这些药物的耐药性决定了广泛耐药结核病。我们评估了一种基于试剂盒的自动化分子检测方法直接从痰标本中检测耐氟喹诺酮类、氨基糖苷类和异烟肼的结核分枝杆菌的准确性。
我们进行了一项前瞻性诊断准确性研究,以比较该研究性检测方法与中国和韩国有结核病症状的成年人的表型药敏试验及DNA测序结果。进行了Xpert MTB/RIF检测和痰培养。结核分枝杆菌分离株进行了表型药敏试验以及katG、gyrA、gyrB和rrs基因及eis和inhA启动子区域的DNA测序。
在308例结核分枝杆菌培养阳性的参与者中,以表型药敏试验作为参考标准时,该研究性检测方法检测耐药性的敏感性分别为:异烟肼83.3%(95%置信区间[CI],77.1%至88.5%)、氧氟沙星88.4%(95%CI,80.2%至94.1%)、临界浓度为0.5μg/ml的莫西沙星87.6%(95%CI,79.0%至93.7%)、临界浓度为2.0μg/ml的莫西沙星96.2%(95%CI,87.0%至99.5%)、卡那霉素71.4%(95%CI,56.7%至83.4%)、阿米卡星70.7%(95%CI,54.5%至83.9%)。除临界浓度为2.0μg/ml的莫西沙星外(特异性为84.0%[95%CI,78.9%至88.3%]),该检测方法检测表型耐药性的特异性对所有药物均为94.3%或更高。以DNA测序作为参考标准时,该研究性检测方法检测与耐药相关突变的敏感性分别为:异烟肼98.1%(95%CI,94.4%至99.6%)、氟喹诺酮类95.8%(95%CI,89.6%至98.8%)、卡那霉素92.7%(95%CI,80.1%至98.5%)以及阿米卡星96.8%(95%CI,83.3%至99.9%),所有药物的特异性均为99.6%(95%CI,97.9%至100%)或更高。
该研究性检测方法能准确检测与异烟肼、氟喹诺酮类和氨基糖苷类耐药相关的结核分枝杆菌突变,有望作为一种快速即时检验方法,为结核病患者的治疗决策提供指导。(由美国国立卫生研究院国家过敏和传染病研究所及中国科学技术部资助;ClinicalTrials.gov编号,NCT02251327。)