Abstract:
Circular RNAs (circRNAs) have recently gained substantial attention as potential biomarkers in several human diseases, most prominently in cancer. However, the detection and quantification of circRNA biomarkers pose specific challenges owing to their circular nature. In particular, the use of enzymes, such as reverse transcriptases, may lead to impaired quantification, poor interlaboratory reproducibility and even false-positive results. Therefore, the development of methods for accurate quantification of circRNA biomarkers is a high priority. An enzyme-free and digital quantification method, named NanoString nCounter, was recently adapted for circRNA detection. In this review, I describe the advantages of using this technology for circRNA quantification as well as methodological considerations, potential pitfalls and disadvantages.
摘要:
环状RNA(circularRNAs,circRNAs)作为几种人类疾病的潜在生物标志物,最近获得了广泛的关注。在癌症中最突出。然而,circRNA生物标志物的检测和定量由于其循环性质而带来了特定的挑战。特别是,酶的使用,比如逆转录酶,可能导致量化受损,实验室间可重复性差,甚至假阳性结果。因此,准确定量circRNA生物标志物的方法的开发是一个高度优先事项。一种无酶和数字定量方法,名为NanoStringnCounter,最近适用于circRNA检测。在这次审查中,我描述了使用这种技术进行circRNA定量的优势以及方法学考虑,潜在的陷阱和缺点。