Optimized detection of lung IL-6 via enzymatic liquefaction of low respiratory tract samples: application for managing ventilated patients,Analyst

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Optimized detection of lung IL-6 via enzymatic liquefaction of low respiratory tract samples: application for managing ventilated patients
Analyst ( IF 3.6 ) Pub Date : 2021-09-17 , DOI: 10.1039/d1an00763g
Antonio Clemente ₁ , Alejandra Alba-Patiño _{1,

2} , Giulia Santopolo _{1,

2} , Enrique Barón ₁ , Estrella Rojo-Molinero ₃ , Antonio Oliver ₃ , Jon Pérez-Bárcena ₄ , Paz Merino de Cos ₅ , María Aranda ₆ , Alberto Del Castillo ₆ , Antonia Socias ₆ , Marcio Borges _{1,

6} , Roberto de la Rica _{1,

2}

Affiliation

Lung IL-6 is a promising biomarker for predicting respiratory failure during pulmonary infections. This biomarker is found in respiratory samples which need to be liquefied prior to analysis. Traditional liquefying methods use reducing agents such as dithiothreitol (DTT). However, DTT impairs immunodetection and does not liquefy highly viscous samples. We propose an enzymatic method that liquefies samples by means of generating O₂ bubbles with endogenous catalase. Low respiratory tract specimens from 48 mechanically ventilated patients (38 with SARS-CoV-2 infection) were treated with DTT or with the enzymatic method. We used turbidimetry to compare the liquefaction degree and IL-6 was quantified with ELISA. Finally, we used AUC-ROC, time-to-event and principal component analysis to evaluate the association between respiratory compromise or local inflammation and IL-6 determined with both methods. Enzymatically treated samples were better liquefied than those reduced by DTT, which resulted in higher ELISA signals. Lung IL-6 levels obtained with the enzymatic procedure were negatively correlated with the oxygenation index (PaO₂/FiO₂) and the time of mechanical ventilation. The proposed enzymatic liquefaction method improves the sensitivity for lung IL-6 detection in respiratory samples, which increases its predictive power as a biomarker for evaluating respiratory compliance.

中文翻译：

通过酶解下呼吸道样本优化肺 IL-6 检测：用于管理通气患者

肺 IL-6 是一种很有前景的生物标志物，可用于预测肺部感染期间的呼吸衰竭。这种生物标志物存在于需要在分析前液化的呼吸道样本中。传统的液化方法使用还原剂，如二硫苏糖醇 (DTT)。然而，DTT 会损害免疫检测，并且不会液化高粘性样品。我们提出了一种通过产生 O ₂来液化样品的酶促方法含有内源性过氧化氢酶的气泡。来自 48 名机械通气患者（38 名感染 SARS-CoV-2）的下呼吸道标本用 DTT 或酶法治疗。我们用浊度法比较液化程度，用ELISA定量IL-6。最后，我们使用 AUC-ROC、事件时间和主成分分析来评估呼吸损害或局部炎症与两种方法测定的 IL-6 之间的关联。酶处理的样品比 DTT 减少的样品液化得更好，这导致了更高的 ELISA 信号。用酶促方法获得的肺 IL-6 水平与氧合指数 (PaO ₂ /FiO ₂)和机械通气时间。所提出的酶促液化方法提高了呼吸道样本中肺 IL-6 检测的灵敏度，从而提高了其作为评估呼吸顺应性的生物标志物的预测能力。

更新日期：2021-09-28

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