College of Medical, Veterinary and Life Sciences, University of Glasgow, Room 202, Sir James Black Building, Glasgow, G128QQ, UK.
Division of Biochemical Diseases, Department of Paediatrics, School of Medicine, BC Children's Hospital, University of British Columbia, Vancouver, BC, V6H 3N1, Canada.
Mol Diagn Ther. 2022 Jan;26(1):19-37. doi: 10.1007/s40291-021-00570-2. Epub 2021 Dec 26.
To evaluate the application of RNAscope in the clinical diagnostic field compared to the current 'gold standard' methods employed for testing gene expression levels, including immunohistochemistry (IHC), quantitative real time PCR (qPCR), and quantitative reverse transcriptase PCR (qRT-PCR), and to detect genes, including DNA in situ hybridisation (DNA ISH).
This systematic review searched CINAHL, Medline, Embase and Web of Science databases for studies that were conducted after 2012 and that compared RNAscope with one or more of the 'gold standard' techniques in human samples. QUADAS-2 test was used for the evaluation of the articles' risk of bias. The results were reviewed narratively and analysed qualitatively.
A total of 27 articles (all retrospective studies) were obtained and reviewed. The 27 articles showed a range of low to middle risk of bias scores, as assessed by QUADAS-2 test. 26 articles studied RNAscope within cancer samples. RNAscope was compared to different techniques throughout the included studies (IHC, qPCR, qRT-PCR and DNA ISH). The results confirmed that RNAscope is a highly sensitive and specific method that has a high concordance rate (CR) with qPCR, qRT-PCR, and DNA ISH (81.8-100%). However, the CR with IHC was lower than expected (58.7-95.3%), which is mostly due to the different products that each technique measures (RNA vs. protein).
This is the first systematic review to be conducted on the use of RNAscope in the clinical diagnostic field. RNAscope was found to be a reliable and robust method that could complement gold standard techniques currently used in clinical diagnostics to measure gene expression levels or for gene detection. However, there were not enough data to suggest that RNAscope could stand alone in the clinical diagnostic setting, indicating further prospective studies to validate diagnostic accuracy values, in keeping with relevant regulations, followed by cost evaluation are required.
评估 RNAscope 在临床诊断领域的应用，与当前用于检测基因表达水平的“金标准”方法（包括免疫组织化学（IHC）、实时定量 PCR（qPCR）和实时定量逆转录 PCR（qRT-PCR））以及用于检测基因的原位杂交技术（DNA ISH）进行比较。
本系统评价检索了 CINAHL、Medline、Embase 和 Web of Science 数据库中 2012 年后发表的研究，比较了 RNAscope 与人类样本中一种或多种“金标准”技术的研究。使用 QUADAS-2 试验评估文章的偏倚风险。结果进行了叙述性综述和定性分析。
共获得并审查了 27 篇文章（均为回顾性研究）。QUADAS-2 试验评估的 27 篇文章显示出低至中等的偏倚风险评分。26 篇文章研究了癌症样本中的 RNAscope。在纳入的研究中，RNAscope 与不同的技术进行了比较（IHC、qPCR、qRT-PCR 和 DNA ISH）。结果证实 RNAscope 是一种高度敏感和特异的方法，与 qPCR、qRT-PCR 和 DNA ISH 具有高一致性率（81.8-100%）。然而，与 IHC 的一致性率低于预期（58.7-95.3%），这主要是由于每种技术测量的产品不同（RNA 与蛋白质）。
这是首次对 RNAscope 在临床诊断领域的应用进行的系统评价。研究发现，RNAscope 是一种可靠且强大的方法，可以补充目前用于临床诊断的金标准技术，以测量基因表达水平或用于基因检测。然而，没有足够的数据表明 RNAscope 可以单独在临床诊断环境中使用，这表明需要进一步的前瞻性研究来验证诊断准确性值，符合相关法规，并随后进行成本评估。
