Elizabeth Herring, Éric Tremblay, Jean-François Beaulieu, Laboratory of Intestinal Physiopathology, Department of Anatomy and Cell Biology, Faculty of Medicine and Health Sciences, Université de Sherbrooke, Sherbrooke, QC J1H5N4, Canada.
World J Gastroenterol. 2017 Apr 28;23(16):2891-2898. doi: 10.3748/wjg.v23.i16.2891.
To investigate the use of droplet digital polymerase chain reaction (ddPCR) for detecting host mRNA markers in stools as a non-invasive test for colorectal cancer screening.
ddPCR and quantitative PCR were compared side by side for their performance in the detection of and transcripts in stool samples obtained from patients with various types of colorectal lesions (advanced adenomas and stage II-IV colorectal cancers) and control (patients displaying no pathological findings) using duplex TaqMan reactions for both methods. and were chosen for this proof-of-concept study based on their relative medium and low abundance in stool samples, respectively, as established in a previous study.
We found that the ddPCR and qPCR methods performed equally well in this TaqMan duplex assay for the detection of and transcripts in stools of patients with colorectal lesions. For , receiver operating characteristic (ROC) curve analysis showed comparable areas under the curve of 0.91 ( < 0.0001) and 0.89-0.90 ( < 0.0001) for the prediction of advanced adenomas and colorectal cancers, respectively. , which was detected at very low levels in control patients, was found to be significantly elevated (over 40 times) in stage II and III colorectal cancers ( < 0.0002). Comparison of the two sets of data revealed a strong correlation of the copy numbers obtained by ddPCR and qPCR for both and .
We found that and detection in stools of patients with colorectal cancers with ddPCR is comparable to that of qPCR using TaqMan assays.
研究利用液滴式数字聚合酶链反应（ddPCR）检测粪便中的宿主 mRNA 标志物作为结直肠癌筛查的非侵入性检测方法。
ddPCR 和定量 PCR 并排比较，用于检测来自患有各种结直肠病变（高级腺瘤和 II-IV 期结直肠癌）和对照组（无病理发现的患者）患者粪便样本中 和 转录本，两种方法均使用双 TaqMan 反应。选择 和 进行此概念验证研究，是基于它们在先前研究中分别在粪便样本中的相对中低丰度。
我们发现，ddPCR 和 qPCR 方法在这种 TaqMan 双探针测定法中，在结直肠病变患者的粪便中检测 和 转录本的性能相当。对于 ，接受者操作特征（ROC）曲线分析显示，用于预测高级腺瘤和结直肠癌的曲线下面积（AUC）分别为 0.91（<0.0001）和 0.89-0.90（<0.0001），具有可比性。在对照组患者中检测到的非常低水平的 ，在 II 期和 III 期结直肠癌中显著升高（超过 40 倍）（<0.0002）。两组数据的比较显示，ddPCR 和 qPCR 获得的拷贝数之间存在很强的相关性。
我们发现，ddPCR 检测结直肠癌患者粪便中的 和 与 TaqMan 测定的 qPCR 相当。
