One-Step In Situ RNA Integrity Analysis for FFPE Tissue

描述(由申请人提供)：将开发一种快速而简单的原位检测福尔马林固定石蜡包埋(FFPE)中RNA完整性的方法，以便于选择具有良好注释的、具有高质量RNA的FFPE临床样本进行基因表达分析。不同长度的猝灭自动连接(QUAL)探针的荧光信号比率将被用来评估在繁琐和耗时的RNA提取之前原位FFPE样品中RNA的裂解程度。QUAL探针的质量控制(QC)指标将与通过RT-PCR和微阵列对从FFPE样本提取的RNA进行RNA完整性分析的下游基因表达分析的质量相关。最终目标是开发并商业化FFPE样本RNA质量的质量控制试剂盒。这项研究将有助于和加速发现更准确的分子生物标记物，用于癌症的检测、预后和治疗。 公共卫生相关性：“FFPE组织的一步原位RNA完整性分析”。世界各地有数百万具有潜在价值的、经过良好注释的临床病理样本，其中许多由于样本质量较差而无法用于临床研究。这项拟议的研究将提高只选择高质量的档案病理样本进行分子分析的准确性，从而使癌症的检测、预后和治疗更加准确。开发一种简单、快速、商业化的检测方法来评估病理样本的质量，也将加快为癌症患者发现药物和在普通人群中检测癌症所需的分子研究的步伐，以改善公众健康。

DESCRIPTION (provided by applicant): A rapid and simple in situ test for RNA integrity in Formalin-Fixed Paraffin Embedded (FFPE) will be developed in order to facilitate the selection of well-annotated, archival FFPE clinical samples with high quality RNA for gene expression analysis. The ratio of fluorescence signal from Quenched Auto Ligation (QUAL) probes of different lengths will be used to assess the degree of RNA fragmentation in FFPE samples in situ, prior to cumbersome and time- consuming RNA extraction. The Quality Control (QC) metric from the QUAL probes will be correlated to the quality of downstream gene expression analysis by RT-PCR and microarray on RNA extracted from the FFPE samples assayed for RNA integrity. The eventual goal is to develop and commercialize a QC kit for FFPE sample RNA quality. This research will aid and accelerate the discovery of more accurate molecular biomarkers for detection, prognosis and treatment of cancer. PUBLIC HEALTH RELEVANCE: "One Step In Situ RNA Integrity Analysis for FFPE Tissue". There are millions of potentially valuable, well-annotated clinical pathology samples around the world many of which cannot be used for clinical studies because of poor sample quality. The proposed research will enable more accurate detection, prognosis and treatment of cancer by improving the accuracy of selecting only high quality archival pathology samples for molecular analysis. The development of a simple and rapid, commercially available test to assess the quality of a pathology sample will also accelerate the pace of molecular studies needed for the discovery of drugs for cancer patients, and the detection of cancer in the general population in order to improve public health.