Sputum Mycobacterium tuberculosis mRNA as a Marker of Bacteriologic Clearance in Response to Antituberculosis Therapy
Sputum Mycobacterium tuberculosis mRNA as a Marker of Bacteriologic Clearance in Response to Antituberculosis Therapy
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DOI：
10.1128/jcm.01526-09
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发表时间：
2010-01-01
影响因子：
9.4
通讯作者：
Eisenach, K. D.
中科院分区：
文献类型：
--
作者：
Li, L.;Mahan, C. S.;Eisenach, K. D.
信使核糖核酸是细胞活力的标志。定量检测痰中结核分枝杆菌mRNA是监测抗结核药物疗效和评价单药疗效的一种很有前景的工具。在一项氟喹诺酮类药物的早期杀菌活性研究中，对接受单一治疗的结核病患者的痰标本中的mRNA水平进行了测量，并在一项白细胞介素2(IL-2)试验中测量了那些接受标准利福平方案的患者的痰标本中的mRNA水平。在早期杀菌活性研究中，于给药前2d和给药后7d每天收集痰进行定量培养和mRNA分析。在IL-2试验中，在整个强化治疗阶段收集痰进行定量培养、Bactec 460液体培养和信使核糖核酸分析。从消化的痰中提取RNA，并用定量逆转录聚合酶链式反应检测几个基因靶点。在接受异烟肼、加替沙星、左氧氟沙星和莫西沙星单一治疗的患者中，乙醛循环酶异柠檬酸裂解酶的mRNA以相似的速度下降。在所有治疗组中，异柠檬酸裂解酶mRNA与治疗前和单一治疗7天期间痰中的CFU高度相关。接受利福平方案治疗1个月的培养阳性结核病患者的痰中可检测到异柠檬酸裂解酶mRNA。在2个月时，液体培养中异柠檬酸mRNA的痰与生长的相关性比固体培养中的更密切。数据表明异柠檬酸裂解酶mRNA是结核分枝杆菌活力的可靠标记物。
mRNA is a marker of cell viability. Quantifying Mycobacterium tuberculosis mRNA in sputum is a promising tool for monitoring response to antituberculosis therapy and evaluating the efficacy of individual drugs. mRNA levels were measured in sputum specimens from patients with tuberculosis (TB) receiving monotherapy in an early bactericidal activity study of fluoroquinolones and in those receiving a standard rifampin-based regimen in an interleukin-2 (IL-2) trial. In the early bactericidal activity study, sputum for quantitative culture and mRNA analysis was collected for 2 days before and daily during 7 days of study drug administration. In the IL-2 trial, sputum was collected for quantitative culture, Bactec 460 liquid culture, and mRNA analysis throughout the intensive treatment phase. RNA was isolated from digested sputum and tested in quantitative reverse transcriptionPCR assays for several gene targets. mRNA for the glyoxylate cycle enzyme isocitrate lyase declined at similar rates in patients receiving isoniazid, gatifloxicin, levofloxacin, and moxifloxacin monotherapy. Isocitrate lyase mRNA correlated highly with CFU in sputum prior to therapy and during 7 days of monotherapy in all treatment arms. Isocitrate lyase mRNA was detectable in sputum of culture-positive TB patients receiving a rifampin-based regimen for 1 month. At 2 months, sputum for isocitrate mRNA correlated more closely with growth in liquid culture than did growth on solid culture medium. Data suggest that isocitrate lyase mRNA is a reliable marker of M. tuberculosis viability.