一种新型、灵敏的LC/MS/MS定量囊性纤维化患者痰液中绿脓杆菌素的方法

A. Ji, D. Rat, Stéphane Muccio, Alexandra Tavernier, L. Taylor, L. L. Chung, S. Richards
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引用次数: 1

摘要

pyoverdine和pyochelin是铜绿假单胞菌在囊性纤维化患者中分泌的铁载体化合物。目前已有的关于囊性纤维化患者痰液中pyoverdine和pyochelin的定量研究文献是基于对检测目标的荧光特性进行检测,而不区分其类型,采用色谱分离的方法。在这些研究中,pyochelin的检出率总是远远低于pyoverdine。这与基因表达数据相反,表明pyochelin荧光检测方法缺乏灵敏度,导致分子不可检测。在本报告中,我们描述了一种新的、灵敏的LC/MS/MS方法,用于定量铜绿假单胞菌(培养阳性)感染的囊性纤维化患者痰中的脓毒杆菌素。使用一种经过验证的生物分析方法,对来自生物库的囊性纤维化患者的26份痰样本进行了分析。数据显示,LC/MS/MS方法符合生物分析验证接受标准,pyochelin检测结果与报道的囊性纤维化患者铜绿假单胞菌浓度检测结果一致。这种定量人痰中脓毒杆菌素的新方法更敏感,可重复性好,更容易可靠地测量囊性纤维化患者的生物标志物脓毒杆菌素。监测囊性纤维化患者痰样品中的pyoverdine和pyochelin可为囊性纤维化新药开发提供更好的分析工具。DOI: 10.29011 / 2576 - 9588.100035
本文章由计算机程序翻译,如有差异,请以英文原文为准。
A Novel and Sensitive LC/MS/MS Method for Quantitation of Pyochelin in Human Sputum Samples from Cystic Fibrosis Patients
Pyoverdines and pyochelin are siderophore compounds secreted by Pseudomonas aeruginosa present in cystic fibrosis affected patients. The available literature on the quantification of pyoverdines and pyochelin in sputa of cystic fibrosis patients is based on detecting the fluorescence properties of the targets detecting pyoverdines and pyochelin without differentiating the types by chromagraphic separation. Within these studies pyochelin was always far less detected than pyoverdines. This is in contrast to gene expression data and suggets the pyochelin fluorescence detection method lacks sensitivity which results in the molecule being non-detectable. In this report, we describe a novel and sensitive LC/MS/MS method for quantitation of pyochelin in human sputum from cystic fibrosis affected patients with Pseudomonas aeruginosa (culture positive) infection. Using a validated bioanalytical method, 26 sputum samples from cystic fibrosis patients from a biobank were analyzed. Data showed the LC/MS/MS method met bioanalytical validation acceptance criteria and results of pyochelin were consistent with reported results of Pseudomonas aeruginosa concentration in cystic fibrosis patients. This new approach for quantitation of pyochelin in human sputum is more sensitive, reproducible and easier to reliably measure the biomarker pyochelin in cystic fibrosis patients. Monitoring pyoverdines and pyochelin in the sputum samples from cystic fibrosis patients may provide better analytical tools for cystic fibrosis new drug development. DOI: 10.29011/2576-9588.100035
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