Department of Biochemistry, Calcutta National Medical College, Kolkata-700014, India.
Clin Chim Acta. 2010 May 2;411(9-10):671-4. doi: 10.1016/j.cca.2010.01.030. Epub 2010 Feb 6.
Cell-free DNA is observed to be more in exudative pleural effusions. Based on this fact development of a clinical chemistry test for classification of pleural effusion will require DNA extraction followed by PCR amplification and electrophoresis. These procedures may not be cost effective for the purpose for classification of pleural effusion as already established parameters are popular for the purpose which can be estimated by comparatively low cost colorimetric procedures. Therefore development of a simple colorimetric test for the classification of pleural fluid based on nucleic acid identification test can be attempted. The aim of this work is to develop such colorimetric test for classification of pleural effusion using only pleural fluid sample.
Cell pellet is obtained from 5 ml pleural fluid which is lysed and subjected to DNA extraction, followed by identification under UV-transilluminator after electrophoresis and orcinol and diphenylamine reaction.
Exudates show extractable DNA from 5 ml biofluid (n=52) which are not observed from transudate (n=32). Orcinol reaction is significantly positive in exudates (n=52) compared to the transudates (n=32). Diphenylamine test cannot differentiate exudate from transudate.
Orcinol reaction of cell lysate obtained from pleural fluid can classify pleural fluid sample into exudate or transudate.
游离 DNA 在渗出性胸腔积液中更为常见。基于这一事实，开发一种用于胸腔积液分类的临床化学测试将需要进行 DNA 提取，然后进行 PCR 扩增和电泳。这些程序对于胸腔积液的分类可能不具有成本效益，因为已经建立的参数已经广泛用于该目的，并且可以通过相对低成本的比色程序进行估计。因此，可以尝试开发一种基于核酸鉴定测试的简单比色测试来分类胸腔积液。这项工作的目的是开发一种仅使用胸腔积液样本即可对胸腔积液进行分类的比色测试。
从 5 毫升胸腔积液中获得细胞沉淀，对其进行裂解，并进行 DNA 提取，然后在电泳后用 UV 透射仪进行鉴定，以及用愈创木酚和二苯胺反应进行鉴定。
渗出液可从 5 毫升生物体液中提取出可提取的 DNA（n=52），而从漏出液中则未观察到（n=32）。与漏出液（n=32）相比，渗出液（n=52）中的愈创木酚反应明显呈阳性。二苯胺试验不能区分渗出液和漏出液。
从胸腔积液中获得的细胞裂解物的愈创木酚反应可以将胸腔积液样本分类为渗出液或漏出液。
