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Comparison of Automated and Manual DNA Isolation Methods of Liquid-Based Cytology Samples

机译：液体基础细胞学样本的自动化和手动DNA分离方法的比较

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Liquid-based cytology (LBC) has been used as a diagnostic tool for cervical cancer for years and is now being adopted for other gynecological cancers. LBC represents an important challenge to ensure that the process yields representative biospecimens for quality control (QC) of diagnostic procedures. In this study, we compare QC parameters (integrity, yield and purity, and polymerase chain reaction [PCR] amplification) of DNA isolated from LBC (N = 296) using two different nucleic acid isolation methods, manual (n = 233) or automated (n = 63). We also evaluated two different types of cytological brushes for sampling from the cervix. Our results suggest that manual isolation (yield 22.81 +/- 1.92 mu g) resulted in increased DNA recovery when compared with automated isolation (yield 9.96 +/- 1.11 mu g) from LBC samples, with a p-value of < 0.0003. We estimated that 98% (53/54) of the samples preserved the integrity of DNA and were suitable for standard molecular biology analyses. The beta-globin gene was amplified in 100% (296/296) of the DNA samples by endpoint PCR. We found no significant difference between the performance of the cytological brushes (p value of < 0.6711) in a general overview. However, when looking at the results from using each brush individually, the manual isolation method was statistically superior to the automated method. Our work illustrates the impact of good QC of preanalytic conditions, which will be important for the application of LBC for developing early detection methods for gynecological cancers.
机译：液体基细胞学（LBC）已被用作宫颈癌的诊断工具多年，现在正在为其他妇科癌症采用。 LBC代表了确保该过程产生代表性生物研究的诊断程序的质量控制（QC）的重要挑战。在本研究中，使用两种不同的核酸隔离方法，手动（n = 233）或自动化（n = 63）。我们还评估了两种不同类型的细胞学刷子，用于从子宫颈取样。我们的研究结果表明，与来自LBC样品的自动分离（产量为9.96 +/-1.11μg）相比，手动隔离（产量22.81 +/-1.92μg）导致DNA恢复增加，P值为<0.0003。我们估计98％（53/54）样品保留了DNA的完整性，适用于标准分子生物学分析。通过终点PCR在100％（296/296）的DNA样品中扩增β-球蛋白基因。我们发现在一般概述中，细胞学刷的性能（P值<0.6711）之间没有显着差异。但是，在单独使用每个刷子的情况下，手动隔离方法统计上优于自动化方法。我们的工作说明了良好QC的QC QC的影响，这对于妇科癌症早期检测方法的应用是重要的。

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