Royal Devon University Healthcare NHS Foundation Trust, Barrack Road, Exeter, EX53BJ, Devon, UK.
Origin Sciences, Granta Park, Cambridge, CB21 6AD, UK.
Tech Coloproctol. 2023 Mar;27(3):227-235. doi: 10.1007/s10151-022-02704-y. Epub 2022 Sep 27.
The ORI-EGI-02 study was designed to test the hypothesis that rectal mucus collected using a novel rectal sampling device (OriCol™), contains sufficient human deoxyribonucleic acid (DNA) of the required quality for Next Generation Sequencing (NGS), for colorectal disease genetic signature discovery.
Using National Institute for Health and Care Research methodology, an internal pilot study was performed in January 2020-May 2021, at four sites in the United Kingdom, to assess the process of recruitment, consent, specimen acquisition and viability for analysis. Following an OriCol test, the sample was stabilized with a buffer solution to preserve the material, which was posted to the laboratory. Samples were processed using QIAamp DNA Blood Midi kit to extract DNA and Quant-iT PicoGreen dsDNA Reagent to quantify the retrieved DNA. DNA integrity was measured by Agilent TapeStation system. 25 ng of human amplifiable DNA was prepared for Next Generation Sequencing (NGS), which was performed on an Illumina NextSeq550 sequencer using the 300-cycle high output kit v2.5.
This study assessed the first 300 patients enrolled to the ORI-EGI-02 Study (n = 800). 290/300 (96.67%) were eligible to undergo OriCol sampling procedure and 285/290 (98.27%) had a successful OriCol sample taken. After transportation, extraction and quantification of DNA, 96.20% (279/290) of the samples had NGS successfully performed for bioinformatic analysis.
Our internal pilot study demonstrated that the OriCol sampling device can capture rectal mucus from unprepared bowel in subjects who could undergo a digital rectal examination. The technique could be applied irrespective of age, frailty, or co-morbidity. Completion of the study to 800 patients and analysis of NGS data for colorectal cancer mutations will now proceed.
ORI-EGI-02 研究旨在检验以下假设，即使用新型直肠取样装置（OriCol™）收集的直肠黏液中含有足够的人类脱氧核糖核酸（DNA），其质量符合下一代测序（NGS）的要求，可用于发现结直肠疾病的遗传特征。
采用英国国家卫生与保健优化研究所的方法，于 2020 年 1 月至 2021 年 5 月在英国的四个地点进行了一项内部试点研究，以评估招募、同意、样本采集和分析可行性的过程。在 OriCol 检测后，使用缓冲液稳定样本以保存材料，然后将样本邮寄到实验室。使用 QIAamp DNA Blood Midi 试剂盒提取 DNA，用 Quant-iT PicoGreen dsDNA 试剂定量检测提取的 DNA。使用 Agilent TapeStation 系统测量 DNA 完整性。准备 25ng 可扩增人类 DNA 进行下一代测序（NGS），在 Illumina NextSeq550 测序仪上使用 300 循环高输出试剂盒 v2.5 进行。
本研究评估了 ORI-EGI-02 研究中纳入的前 300 名患者（n=800）。300 名患者中有 290 名（96.67%）符合接受 OriCol 取样程序的条件，其中 285 名（98.27%）成功进行了 OriCol 取样。经过运输、提取和 DNA 定量后，290 名患者中有 279 名（96.20%）的样本成功进行了 NGS 生物信息分析。
我们的内部试点研究表明，OriCol 取样装置可以从接受数字直肠检查的未经准备的肠道中采集直肠黏液。该技术可以应用于任何年龄、虚弱或合并症患者。目前正在继续进行 800 名患者的研究并分析 NGS 数据以发现结直肠癌突变。
