Application of DNA- and RNA-based sequencing techniques to tumour tissue samples in the clinical laboratory
应用基于DNA和rna的测序技术在临床实验室肿瘤组织样本
Abstract
Next-generation sequencing (NGS) technologies have become an indispensable tool within the research field driving genomic discoveries and furthering our understanding of the genomic changes that lead to human disease. NGS technologies have great potential to provide invaluable genomic data that can be used to improve clinical diagnosis and the delivery of precision medicine. Over the past decade NGS has translated into the clinical setting for both hereditary and somatic indications. Whilst it has great potential in the clinic, the application of this technology for cancer faces a number of challenges, both technical and logistic. Consideration must be given to the role of this technology and how it is best used in the patient pathway. In this review we describe the current technologies routinely used in the clinical laboratory and provide insights into their application in solid tumour testing.
新一代测序(NGS)技术已经成为研究领域中一个不可或缺的工具，推动基因组发现，并进一步了解导致人类疾病的基因组变化。
NGS技术有巨大的潜力提供无价的基因组数据，这些数据可用于改善临床诊断和精准医疗的提供。
在过去的十年中，NGS已经转化为遗传和躯体适应症的临床设置。
虽然它在临床中有巨大的潜力，但这项技术在癌症方面的应用面临着技术和后勤方面的诸多挑战。
必须考虑到这项技术的作用，以及它如何最好地应用于患者路径。
在这篇综述中，我们描述了目前在临床实验室常规使用的技术，并提供了他们在实体肿瘤检测中的应用见解。
Keywords
Introduction
Over the past decade next-generation sequencing (NGS) has emerged as a vital tool in the clinical diagnostic setting given its ability to determine the identity and order of nucleotides in a DNA or RNA molecule (DNA-seq/RNA-seq). Next generation sequencing describes the emergence of non-Sanger based sequencing technologies, which enable a substantially increased sequencing throughput by enabling millions of targets to be sequenced in parallel, hence the commonly applied description of ‘massively parallel sequencing’.
The transition of NGS technologies into the clinical laboratory has progressed significantly since the introduction of the first commercially available next generation sequencer in 2004. Healthcare laboratories across the world have looked to harness the power of NGS in order to provide improved clinical outcomes for patients. NGS in the clinical setting has been led by constitutional genetic testing, where expensive, time consuming sequential testing has been replaced by simultaneous gene panel analysis. The challenges faced by constitutional testing and solid tumour testing (Table 1) share many similarities, however a number of additional challenges face solid tumour testing and its uptake into the clinical laboratory.1
在过去的十年中，下一代测序(NGS)已经成为临床诊断环境中的一个重要工具，因为它能够确定DNA或RNA分子(DNA-seq/RNA-seq)中核苷酸的身份和顺序。
下一代测序技术描述了基于非桑格序列的测序技术的出现，这种技术通过使数百万个靶点并行测序，从而大大提高了测序通量，因此通常被称为“大规模并行测序”。
NGS技术的过渡到临床实验室已经取得显著进展，自从引进第一个商业可获得的下一代测序器在2004年。
全世界的医疗保健实验室都在寻求利用NGS的力量，以便为患者提供更好的临床结果。
在临床环境下的NGS已经被结构基因测试所主导，其中昂贵的，耗时的序列测试已经被同时的基因面板分析所取代。
体质检测和实体肿瘤检测(表1)所面临的挑战有许多相似之处，但是实体肿瘤检测和临床实验室的应用还面临一些额外的挑战。