Purpose: For anaplastic lymphoma kinase (ALK) gene detection, the centrifugal sedimentation method (CSM) and cell block method (CBM) are commonly used to process samples of bloody pleural effusions (BPEs). However, in practice, the impurity content in the processed samples often affects the results and even leads to the detection failure. The purpose of this study was to establish a cell enrichment method (CEM) by using a disposable membrane cell collector to remove blood and inflammatory cells and enrich lung adenocarcinoma cells in BPE for more efficient RNA extraction and ALK gene detection. Materials and Methods: CEM proposed in this study and the traditional CSM and CBM were used to treat BPE samples collected from 37 lung adenocarcinoma patients. A DeNovix DS-11 ultraviolet spectrophotometer was used to measure the concentration and purity of extracted RNA. Amplification refractory mutation systems (ARMS) and ABI 7500 fluorescence qPCR were used to detect ALK gene. Through statistical analysis, the CEM was compared with the CSM and CBM in RNA concentration, purity, and ALK gene detection results. Results: The concentration of RNA extracted by using the CEM was significantly higher than that extracted by using the CBM and CSM (p #x3c; 0.001). The purity of RNA extracted by using the CEM was significantly higher than that by the other 2 methods (p = 0.011, p = 0.005). ALK gene testing with PCR was successful in all the samples using the CEM, but 2 cases by the CSM and 1 case by the CBM failed. Conclusions: Using the disposable membrane cell collector to process BPE of lung adenocarcinoma patients for RNA extraction and ALK gene detection is more effective and successful compared with the traditional methods, and it is suggested to be further applied and popularized in clinical practice.
Acta Cytologica

中文翻译：

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使用一次性膜细胞收集器富集血性胸腔积液中的肺腺癌细胞，用于间变性淋巴瘤激酶融合基因检测。

目的：为了检测间变性淋巴瘤激酶（ALK）基因，通常使用离心沉降法（CSM）和细胞阻断法（CBM）处理血性胸腔积液（BPE）样品。但是，实际上，处理后的样品中的杂质含量通常会影响结果，甚至导致检测失败。这项研究的目的是建立一种细胞富集方法（CEM），方法是使用一次性膜细胞收集器去除血液和炎性细胞，并富集BPE中的肺腺癌细胞，以更有效地提取RNA和检测ALK基因。材料和方法：这项研究中提出的CEM和传统的CSM和CBM用于处理从37例肺腺癌患者中收集的BPE样品。使用DeNovix DS-11紫外分光光度计测量提取的RNA的浓度和纯度。扩增难治性突变系统（ARMS）和ABI 7500荧光定量PCR用于检测ALK基因。通过统计分析，将CEM与CSM和CBM的RNA浓度，纯度和ALK基因检测结果进行了比较。结果：使用CEM提取的RNA浓度显着高于使用CBM和CSM提取的RNA（p＃x3c; 0.001）。使用CEM提取的RNA的纯度显着高于其他两种方法（p = 0.011，p = 0.005）。使用CEM在所有样品中通过PCR进行的ALK基因测试均成功，但是CSM有2例，CBM有1例失败。结论：与传统方法相比，使用一次性膜细胞收集器处理肺腺癌患者的BPE进行RNA提取和ALK基因检测更为有效和成功，建议在临床上进一步推广应用。
细胞学学报