Methodology for Sputum Induction and Laboratory Processing
Methodology for Sputum Induction and Laboratory Processing
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DOI：
10.3791/56612
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发表时间：
2017-12-01
影响因子：
1.2
通讯作者：
Moermans, Catherine
中科院分区：
文献类型：
--
作者：
Guiot, Julien;Demarche, Sophie;Moermans, Catherine
痰液诱导和处理技术是一种公认的非侵入性方法，允许从气道收集和分析细胞，这在各种呼吸系统疾病如哮喘、慢性阻塞性肺病（COPD）、慢性咳嗽或特发性肺纤维化中是令人感兴趣的。这种技术耐受性好，安全且无创，但目前仅限于临床实践中的研究服务和专业中心，因为它技术要求高，耗时，并且需要训练有素的工作人员。痰液诱导和分析的成功率约为80%，现就痰液样本的诱导和实验室处理进行介绍。通过吸入高渗或等渗盐水和沙丁胺醇诱导痰。对于处理，我们使用全痰技术。二硫苏糖醇（DTT）用于痰液样本的粘液溶解。痰液处理的主要目的是获得细胞分类计数，以研究气道腔中存在的细胞类型。还可以对痰液上清液和痰液细胞进行额外的分析，这可以进一步研究炎症过程和免疫机制。例如研究痰液上清液中的介质，对痰液细胞进行大范围的分析，如流式细胞术、基因组学或蛋白质组学。最后，给出了健康对照、哮喘和COPD患者痰液分析的代表性结果。
The technique of sputum induction and processing is a recognized non-invasive method allowing the collection and analysis of cells from the airways, which is interesting in various respiratory diseases like asthma, chronic obstructive pulmonary disease (COPD), chronic cough, or idiopathic pulmonary fibrosis. This technique is well tolerated, safe and non-invasive, but is currently limited to research services and specialized centers in clinical practice because it is technically demanding, time-consuming, and requires trained staff. The success rate of sputum induction and analysis is about 80%.Here, we describe the induction and laboratory processing of sputum samples. Sputum is induced by inhalation of hypertonic or isotonic saline with salbutamol. For the processing, we use the whole sputum technique. Dithiothreitol (DTT) is used to allow mucolysis of sputum samples. The primary aim of sputum processing is to obtain a differential cell count to study the cell types present in the airway lumen. Additional analyses may also be performed on sputum supernatant and sputum cells, which may allow further investigation into inflammatory processes and immune mechanisms. Examples include studying mediators in sputum supernatant and performing a large spectrum of analysis on sputum cells such as flow cytometry, genomics, or proteomics.Finally, representative results of sputum analysis in healthy controls, asthmatics, and COPD patients are presented.