Optimizing RNA Extraction of Renal Papilla Biopsy Tissue in Kidney Stone Formers: A New Methodology for Genomic Study.
Optimizing RNA Extraction of Renal Papilla Biopsy Tissue in Kidney Stone Formers: A New Methodology for Genomic Study.
复制标题
优化肾结石形成者肾乳头活检组织的 RNA 提取：基因组研究的新方法。
作者：
Taguchi K;Usawachintachit M;Hamamoto S;Unno R;Tzou DT;Sherer BA;Wang Y;Okada A;Stoller ML;Yasui T;Chi T.
简介：内窥镜工具为肾结石手术提供了多种检查和治疗。尽管腔内泌尿外科医生使用内窥镜收集组织来研究泌尿结石疾病，但这种组织收集方法是有限的。内窥镜下切除的组织尺寸较小，限制了可能的基于基因组的检查类型。我们研究了一种新的方法，肾乳头活检和RNA提取，以建立一个基因组研究方法为肾结石diseases.Patients和方法：我们进行了一项前瞻性的多机构研究，并收集肾乳头标本连续经皮肾镜取石术和输尿管镜（URS）的情况下进行去除上尿路结石。取石后用输尿管镜活检钳取肾乳头组织。使用两种不同的提取试剂盒提取RNA，并检查其数量和质量。此外，活检对手术并发症的影响进行了比较，与不进行活检的情况下，匹配的病例对照分析调整年龄，性别，体重指数，bilaterality，和石burn.Results：共90活检49例患者进行，标本采集和RNA提取之间的中位持续时间为61天。单变量和多变量分析均显示，BIGopsy®镊子的使用显著增加了提取RNA的总产量（p= 0.004）和质量（A260/280 p= 0.001，A260/A230 p= 0.004）。使用RNeasy Micro Kit®的提取也改善了A260/A230，而通过单变量和多变量分析降低了提取的RNA的RNA完整性数量（分别为p= 0.002和p < 0.001）。此外，配对病例对照研究表明，内窥镜肾乳头活检术未引起明显的手术并发症，包括出血、结石清除率和红细胞压积下降以及肾功能不全。在URS的活检给予平均20分钟的程序时间在nonbiopsy cases.Conclusions：我们证明了一种安全的方法，最佳的RNA提取肾乳头组织。这项技术将通过促进更大的组织产量来加速肾结石形成者的先进基因组研究。
Introduction:Endoscopic tools have provided versatile examination and treatment for kidney stone procedures. Despite endourologists researching urinary stone disease using endoscopes to collect tissue, this tissue collection method is limited. Endoscopically removed tissues are small in size, restricting the types of genome-based examination possible. We investigated a new method of renal papilla biopsy and RNA extraction to establish a genomic research methodology for kidney stone disease.Patients and Methods:We conducted a prospective multi-institutional study and collected renal papilla specimens from consecutive percutaneous nephrolithotomy and ureteroscopy (URS) cases performed for removal of upper urinary tract stones. Renal papilla tissue was extracted using ureteroscopic biopsy forceps after stone removal. RNA was extracted using two different extraction kits, and their quantity and quality were examined. Additionally, the impact of biopsy on surgical complications was compared between cases performed with and without biopsy by matched case–control analysis adjusted for age, gender, body mass index, bilaterality, and stone burden.Results:A total of 90 biopsies from 49 patients were performed, and the median duration between specimen collection and RNA extraction was 61 days. Both univariate and multivariate analyses showed BIGopsy®forceps usage significantly increased the total yield (p= 0.004) and quality (p= 0.001 for A260/280,p= 0.004 for A260/A230) of extracted RNA. Extraction using the RNeasy Micro Kit®also improved A260/A230, whereas reduced RNA integrity number of extracted RNA by univariate and multivariate analyses (p= 0.002 andp< 0.001, respectively). Moreover, matched case–control study demonstrated that endoscopic renal papilla biopsy caused no significant surgical complications, including bleeding, decreased stone clearance and hematocrit, and renal dysfunction. Biopsies during URS imparted an average of 20 minutes of procedure time over nonbiopsy cases.Conclusions:We demonstrate a safe methodology for optimal RNA extraction of renal papilla tissue. This technique will accelerate advanced genomic studies for kidney stone formers by facilitating larger tissue yields.