Berna Amalia Z, DeBosch Brian, Stoll Janis, Odom John Audrey R
Department of Pediatrics, Washington University School of Medicine.
Department of Pediatrics, Washington University School of Medicine; Cell Biology & Physiology, Washington University School of Medicine.
J Vis Exp. 2019 Feb 14(144). doi: 10.3791/59217.
Breath collection and analysis can be used to discover volatile biomarkers in a number of infectious and non-infectious diseases, such as malaria, tuberculosis, lung cancer, and liver disease. This protocol describes a reproducible method for sampling breath in children and then stabilizing breath samples for further analysis with gas chromatography-mass spectrometry (GC-MS). The goal of this method is to establish a standardized protocol for the acquisition of breath samples for further chemical analysis, from children aged 4-15 years. First, breath is sampled using a cardboard mouthpiece attached to a 2-way valve, which is connected to a 3 L bag. Breath analytes are then transferred to a thermal desorption tube and stored at 4-5 °C until analysis. This technique has been previously used to capture breath of children with malaria for successful breath biomarker identification. Subsequently, we have successfully applied this technique to additional pediatric cohorts. The advantage of this method is that it requires minimal cooperation on part of the patient (of particular value in pediatric populations), has a short collection period, does not require trained staff, and can be performed with portable equipment in resource-limited field settings.
呼吸采集与分析可用于发现多种感染性和非感染性疾病中的挥发性生物标志物,如疟疾、肺结核、肺癌和肝病。本方案描述了一种可重复的方法,用于采集儿童的呼吸样本,然后稳定呼吸样本,以便用气相色谱 - 质谱联用仪(GC-MS)进行进一步分析。该方法的目标是为4至15岁儿童建立一个标准化的呼吸样本采集方案,用于进一步的化学分析。首先,使用连接到双向阀的硬纸板吹嘴采集呼吸样本,双向阀连接到一个3升的袋子。然后将呼吸分析物转移到热解吸管中,并在4至5°C下储存直至分析。该技术先前已用于采集疟疾患儿的呼吸样本,成功鉴定出呼吸生物标志物。随后,我们已成功将该技术应用于其他儿科队列。该方法的优点是患者所需的配合极少(在儿科人群中具有特殊价值),采集时间短,不需要训练有素的工作人员,并且可以在资源有限的现场环境中使用便携式设备进行操作。