Liquid Chromatography Mass Spectrometry Detection of Antibiotic Agents in Sputum from Persons with Cystic Fibrosis.
Liquid Chromatography Mass Spectrometry Detection of Antibiotic Agents in Sputum from Persons with Cystic Fibrosis.
复制标题
液相色谱质谱法检测囊性纤维化患者痰液中的抗生素。
DOI：
10.1128/aac.00927-20
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发表时间：
2021
影响因子：
4.9
通讯作者：
Whiteson,KatrineL
中科院分区：
文献类型：
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作者：
Gallagher,Tara;Riedel,Stefan;Kapcia,Joseph;Caverly,LindsayJ;Carmody,Lisa;Kalikin,LindaM;Lu,Junnan;Phan,Joann;Gargus,Matthew;Kagawa,Miki;Leemans,SimonW;Rothman,JasonA;Grun,Felix;LiPuma,JohnJ;Whiteson,KatrineL
抗生素治疗预计会对宿主微生物群落产生相当大的影响，然而，许多关注微生物组和健康的研究往往受到抗生素暴露信息有限的困扰。鉴于抗生素具有不同的药代动力学和抗菌特性，研究这些药物在特定宿主标本中的类型和浓度将为了解它们对其中微生物的影响提供急需的见解。在这里，我们建立了液相色谱-质谱（LC-MS）方法来检测囊性纤维化患者痰液中的18种抗生素。采用抗生素峰值对照样品对Waters Acquity Quattro Premier XE的三种液体提取方法进行比较。二硫苏糖醇提取法捕获了最多的抗生素，并用于检测11名囊性纤维化患者的痰样本中的抗生素，并将结果与个体自我报告的抗生素使用情况进行比较。痰液样品采用两次LC-MS分析；Quattro Premier检测到16种抗生素的纳摩尔或微摩尔浓度，而Xevo TQ-XS检测到所有18种抗生素，大多数在亚纳摩尔水平。在45%的痰样本（71/158）中，两种LC-MS方法都检测到至少一种受试者未报告的抗生素，这种不一致在很大程度上是由于阿奇霉素每周给药三次和半衰期长。对于约37%的样品，两种仪器均未检测到报告由个体服用的抗生素。我们的研究结果提供了一种在感染部位检测各种抗生素的方法，从而提供了一种将抗生素使用数据纳入微生物组研究的方法。
Antibiotic therapy is expected to impact host microbial communities considerably, yet many studies focused on microbiome and health are often confounded by limited information about antibiotic exposure. Given that antibiotics have diverse pharmacokinetic and antimicrobial properties, investigating the type and concentration of these agents in specific host specimens would provide much needed insight into their impact on the microbes therein. Here, we developed liquid chromatography mass spectrometry (LC-MS) methods to detect 18 antibiotic agents in sputum from persons with cystic fibrosis. Antibiotic spike-in control samples were used to compare three liquid extraction methods on the Waters Acquity Quattro Premier XE. Extraction with dithiothreitol captured the most antibiotics and was used to detect antibiotics in sputum samples from 11 people with cystic fibrosis, with results being compared to the individuals’ self-reported antibiotic use. For the sputum samples, two LC-MS assays were used; the Quattro Premier detected nanomolar or micromolar concentrations of 16 antibiotics, whereas the Xevo TQ-XS detected all 18 antibiotics, most at subnanomolar levels. In 45% of tested sputum samples (71/158), at least one antibiotic that was not reported by the subject was detected by both LC-MS methods, a discordance largely explained by the thrice weekly administration and long half-life of azithromycin. For ∼37% of samples, antibiotics reported as taken by the individual were not detected by either instrument. Our results provide an approach for detecting a variety of antibiotics at the site of infection, thereby providing a means to include antibiotic usage data into microbiome studies.