Table_1_Accurate isolation and detection of circulating tumor cells using enrichment-free multiparametric high resolution imaging.xlsx

IntroductionThe reliable and accurate detection of rare circulating tumor cells (CTCs) from cancer patient blood samples promises advantages in both research and clinical applications. Numerous CTC detection methods have been explored that rely on either the physical properties of CTCs such as density, size, charge, and/or their antigen expression profiles. Multiple factors can influence CTC recovery including blood processing method and time to processing. This study aimed to examine the accuracy and sensitivity of an enrichment-free method of isolating leukocytes (AccuCyte® system) followed by immunofluorescence staining and high-resolution imaging (CyteFinder® instrument) to detect CTCs.MethodHealthy human blood samples, spiked with cancer cells from cancer cell lines, as well as blood samples obtained from 4 subjects diagnosed with cancer (2 pancreatic, 1 thyroid, and 1 small cell lung) were processed using the AccuCyte-CyteFinder system to assess recovery rate, accuracy, and reliability over a range of processing times.ResultsThe AccuCyte-CyteFinder system was highly accurate (95.0%) at identifying cancer cells in spiked-in samples (in 7.5 mL of blood), even at low spiked-in numbers of 5 cells with high sensitivity (90%). The AccuCyte-CyteFinder recovery rate (90.9%) was significantly higher compared to recovery rates obtained by density gradient centrifugation (20.0%) and red blood cell lysis (52.0%). Reliable and comparable recovery was observed in spiked-in samples and in clinical blood samples processed up to 72 hours post-collection. Reviewer analysis of images from spiked-in and clinical samples resulted in high concordance (R-squared value of 0.998 and 0.984 respectively).DiscussionThe AccuCyte-CyteFinder system is as an accurate, sensitive, and clinically practical method to detect and enumerate cancer cells. This system addresses some of the practical logistical challenges in incorporating CTCs as part of routine clinical care. This could facilitate the clinical use of CTCs in guiding precision, personalized medicine.

引言：对癌症患者血液样本中稀有循环肿瘤细胞（CTCs）的可靠且精确检测，在科研及临床应用方面均具有显著优势。众多CTCs检测方法已探索，这些方法或依赖于CTCs的物理特性，如密度、尺寸、电荷及其抗原表达谱。多种因素可影响CTCs的回收，包括血液处理方法和处理时间。本研究旨在评估一种无需富集的分离白细胞方法（AccuCyte®系统）的准确性和灵敏度，该方法随后通过免疫荧光染色和高清成像（CyteFinder®仪器）来检测CTCs。方法：采用AccuCyte-CyteFinder系统处理健康人血液样本，其中掺入癌细胞系中的癌细胞，以及从4名癌症患者（2名胰腺癌、1名甲状腺癌和1名小细胞肺癌）获取的血液样本，以评估在不同处理时间下的回收率、准确性和可靠性。结果：AccuCyte-CyteFinder系统在识别掺入样本中的癌细胞方面高度准确（95.0%），即使在低掺入数量（5个细胞）时也表现出高灵敏度（90%）。AccuCyte-CyteFinder的回收率（90.9%）显著高于密度梯度离心（20.0%）和红细胞溶解（52.0%）获得的回收率。在掺入样本和临床血液样本中观察到可靠的、可比较的回收，且样本处理时间可延长至采集后72小时。审稿人对掺入样本和临床样本图像的分析结果显示高度一致性（R平方值分别为0.998和0.984）。讨论：AccuCyte-CyteFinder系统是一种准确、灵敏且临床实用的检测和计数癌细胞的方法。该系统解决了将CTCs纳入常规临床护理中的一些实际物流挑战，这可能有助于指导精准、个性化的医疗中CTCs的临床应用。