Bennett William E, González-Rivera Rosbel, Shaikh Nurmohammad, Magrini Vincent, Boykin Michelle, Warner Barbara B, Hamvas Aaron, Tarr Phillip I
Washington University School of Medicine, Department of Pediatrics, Division of Pediatric Gastroenterology, Washington, United States of America.
J Immunol Methods. 2009 Sep 30;349(1-2):56-60. doi: 10.1016/j.jim.2009.07.013. Epub 2009 Aug 3.
Efforts to characterize the human transcriptome have largely been limited to blood, urine, and tissue analyses (i.e., normally sterile materials). We report here an extraction protocol using commercially available reagents to obtain high-yield, reverse-transcribable RNA from human stool. Quantitative reverse transcriptase polymerase chain reactions demonstrated minimal intra-specimen but considerable intra-subject variability over time of transcripts for interleukin-6 (IL-6), IL-8, epidermal growth factor (EGF), calprotectin, and glyceraldehyde-3-phosphate dehydrogenase (GAPDH). This technique now expands opportunities to use the human fecal transcriptome to characterize gastrointestinal pathophysiology.
对人类转录组进行特征描述的工作在很大程度上局限于血液、尿液和组织分析(即通常无菌的材料)。我们在此报告一种提取方案,该方案使用市售试剂从人类粪便中获得高产量、可用于逆转录的RNA。定量逆转录聚合酶链反应表明,白细胞介素-6(IL-6)、IL-8、表皮生长因子(EGF)、钙卫蛋白和甘油醛-3-磷酸脱氢酶(GAPDH)转录本在样本内的差异最小,但在个体内随时间存在相当大的变异性。这项技术现在为利用人类粪便转录组来描述胃肠道病理生理学提供了更多机会。