Altered exhaled biomarker profiles inchildren during and after rhinovirusinduced wheeze
电子鼻在鼻病毒引起的喘息期间和后期改变儿童的呼出生物标志物图谱研究
Marc P. van der Schee1,2,3, Simone Hashimoto1, Annemarie C. Schuurman1,
Janine S. Repelaer van Driel1, Nora Adriaens1, Romy M. van Amelsfoort1,
Tessa Snoeren1, Martine Regenboog1, Aline B. Sprikkelman2, Eric G. Haarman3,
Wim M.C. van Aalderen2 and Peter J. Sterk1
Affiliations:1Dept of Respiratory Medicine, Academic Medical Centre, University of Amsterdam, Amsterdam,
The Netherlands. 2Dept of Pediatric Respiratory Medicine and Allergy, Emma’s Children Hospital, Academic
Medical Centre, University of Amsterdam, Amsterdam, The Netherlands. 3Dept of Pediatric Respiratory
Medicine, VU Medical Centre, VU University of Amsterdam, Amsterdam, The Netherlands.
Correspondence: M.P. van der Schee, Dept of Respiratory Medicine, F5-158, Academic Medical Centre,
University of Amsterdam, Meibergdreef 9, 1105 AZ Amsterdam, The Netherlands.
: m.p.vanderschee@amc.uva.nl
Abstract
Preschool rhinovirus-induced wheeze is associated with an increased risk of asthma. In adult asthma, exhaled volatile organic compounds (VOC) are associated with inflammatory activity. We therefore hypothesised that acute preschool wheeze is accompanied by a differential profile of exhaled VOC, which is maintained after resolution of symptoms in those children with rhinovirus-induced wheeze.
We included 178 children (mean±sd age 22±9 months) from the EUROPA cohort comparing asymptomatic and wheezing children during respiratory symptoms and after recovery. Naso- and oropharyngeal swabs were tested for rhinovirus by quantitative PCR. Breath was collected via a spacer and analysed using an electronic nose. Between-group discrimination was assessed by constructing a 1000-fold cross-validated receiver operating characteristic curve. Analyses were stratified by rhinovirus presence/absence.
Wheezing children demonstrated a different VOC profile when compared with asymptomatic children (p<0.001), regardless of the presence (area under the curve (AUC) 0.77, 95% CI 0.07) or absence (AUC 0.81, 95% CI 0.05) of rhinovirus. After symptomatic recovery, discriminative accuracy was maintained in children with rhinovirus-induced wheeze (AUC 0.84, 95% CI 0.06), whereas it dropped significantly in infants with non-rhinovirus-induced wheeze (AUC 0.67, 95% CI 0.06).
Exhaled molecular profiles differ between preschool children with and without acute respiratory wheeze. This appears to be sustained in children with rhinovirus-induced wheeze after resolution of symptoms. Therefore, exhaled VOC may qualify as candidate biomarkers for early signs of asthma.
学龄前鼻病毒引起的喘息与哮喘风险增加有关。在成人哮喘中，呼出的挥发性有机化合物(VOC)与炎症活性相关。因此，我们假设急性学龄前喘息伴随着呼出VOC的差异性特征，这种特征在鼻病毒诱导喘息的儿童症状消失后得以维持。
我们纳入了来自欧洲队列的178名儿童(平均年龄为22±9个月，标准差)，比较了无症状和喘息儿童在呼吸症状期间和康复后的情况。用定量PCR法检测鼻病毒和口*。通过间隔器收集呼吸，并用电子鼻进行分析。通过构建1000倍交叉验证的受试者操作特征曲线来评估组间差异。根据鼻病毒存在/不存在对分析进行分层。
与无症状儿童相比，哮喘儿童表现出不同的VOC特征(p<0.001)，无论鼻病毒的存在(曲线下面积(AUC)为0.77，95%CI为0.07)或不存在(AUC为0.81，95%CI为0.05)。症状恢复后，鼻病毒性哮喘患儿的鉴别准确率保持不变(AUC 0.84，95%CI 0.06)，而非鼻病毒性哮喘患儿的鉴别准确率显著下降(AUC 0.67，95%CI 0.06)。
有或无急性呼吸性喘息的学龄前儿童的呼气分子特征不同。这似乎是持续的儿童鼻病毒诱导的喘息症状解决后。因此，呼出的VOC可作为哮喘早期症状的候选生物标志物。
Breath collection and analysis
Exhaled breath was collected by means of a Babyhaler (GlaxoSmithKline, Brentford, UK) with inverted valve system. Children breathed tidally into a face mask covering the mouth and nose. Environmental VOC inside the child’s airways and the spacer were washed out during 100 s of breathing through an inspiratory VOC filter (North Safety Products, Middelburg, the Netherlands). Ambient air drawn through an identical VOC filter was used to create a baseline reference signal. The exhaled breath was subsequently sampled for 40 s directly from the spacer by two separate Cyranose 320 Electronic Nose (eNose) devices . A Cyranose 320 has an array of 32 nano-sensors that show nonselective interactions with the VOC mixture. This creates a so-called breathprint, a pattern of resistance changes for the individual sensors that reflects the profile of the VOC mixture. This device has commonly been used for breath analysis, and has shown adequate repeatability and reproducibility [24]. Further details on methodology are available in the online supplementary material.
呼出的呼吸是通过带倒置瓣膜系统的Babyhaler(英国葛兰素史克公司)收集的。孩子们整齐地呼吸着和鼻子。通过吸气式VOC过滤器(荷兰米德尔堡North Safety Products，Middelburg)呼吸100 s时，儿童气道内的环境VOC和间隔物被冲掉。环境空气通过一个相同的VOC过滤器被用来创建一个基准参考信号。随后，通过两个单独的Cyranose 320电子鼻(Enose)装置直接从间隔棒上取样40秒。Cyranose320有32个纳米传感器阵列，显示与VOC混合物的非选择性相互作用。这就产生了一种所谓的呼吸印痕，即反映VOC混合物剖面的单个传感器的电阻变化模式。该装置通常用于呼吸分析，并显示出足够的重复性和再现性[24]。有关方法的更多详细信息，请参阅在线补充材料。
