Altered exhaled biomarker profiles inchildren during and after rhinovirusinduced wheeze
電子鼻在鼻病毒引起的喘息期間和后期改變兒童的呼出生物標(biāo)志物圖譜研究
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.
學(xué)齡前鼻病毒引起的喘息與哮喘風(fēng)險增加有關(guān)。在成人哮喘中，呼出的揮發(fā)性有機(jī)化合物(VOC)與炎癥活性相關(guān)。因此，我們假設(shè)急性學(xué)齡前喘息伴隨著呼出VOC的差異性特征，這種特征在鼻病毒誘導(dǎo)喘息的兒童癥狀消失后得以維持。
我們納入了來自歐洲隊列的178名兒童(平均年齡為22±9個月，標(biāo)準(zhǔn)差)，比較了無癥狀和喘息兒童在呼吸癥狀期間和康復(fù)后的情況。用定量PCR法檢測鼻病毒和口*。通過間隔器收集呼吸，并用電子鼻進(jìn)行分析。通過構(gòu)建1000倍交叉驗證的受試者操作特征曲線來評估組間差異。根據(jù)鼻病毒存在/不存在對分析進(jìn)行分層。
與無癥狀兒童相比，哮喘兒童表現(xiàn)出不同的VOC特征(p<0.001)，無論鼻病毒的存在(曲線下面積(AUC)為0.77，95%CI為0.07)或不存在(AUC為0.81，95%CI為0.05)。癥狀恢復(fù)后，鼻病毒性哮喘患兒的鑒別準(zhǔn)確率保持不變(AUC 0.84，95%CI 0.06)，而非鼻病毒性哮喘患兒的鑒別準(zhǔn)確率顯著下降(AUC 0.67，95%CI 0.06)。
有或無急性呼吸性喘息的學(xué)齡前兒童的呼氣分子特征不同。這似乎是持續(xù)的兒童鼻病毒誘導(dǎo)的喘息癥狀解決后。因此，呼出的VOC可作為哮喘早期癥狀的候選生物標(biāo)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.
呼出的呼吸是通過帶倒置瓣膜系統(tǒng)的Babyhaler(英國葛蘭素史克公司)收集的。孩子們整齊地呼吸著和鼻子。通過吸氣式VOC過濾器(荷蘭米德爾堡North Safety Products，Middelburg)呼吸100 s時，兒童氣道內(nèi)的環(huán)境VOC和間隔物被沖掉。環(huán)境空氣通過一個相同的VOC過濾器被用來創(chuàng)建一個基準(zhǔn)參考信號。隨后，通過兩個單獨的Cyranose 320電子鼻(Enose)裝置直接從間隔棒上取樣40秒。Cyranose320有32個納米傳感器陣列，顯示與VOC混合物的非選擇性相互作用。這就產(chǎn)生了一種所謂的呼吸印痕，即反映VOC混合物剖面的單個傳感器的電阻變化模式。該裝置通常用于呼吸分析，并顯示出足夠的重復(fù)性和再現(xiàn)性[24]。有關(guān)方法的更多詳細(xì)信息，請參閱在線補(bǔ)充材料。
