Greenwood Genetic Center, 113 Gregor Mendel Circle, Greenwood, SC 29646, USA.
J Mol Biol. 2013 Nov 1;425(21):3914-8. doi: 10.1016/j.jmb.2013.07.023. Epub 2013 Jul 20.
The completion of the Human Genome Project provided a reference sequence to which researchers could compare sequences from individual patients in the hope of identifying disease-causing mutations. However, this still necessitated candidate gene testing or a very limited screen of multiple genes using Sanger sequencing. With the advent of high-throughput Sanger sequencing, it became possible to screen hundreds of patients for alterations in hundreds of genes. This process was time consuming and limited to a few locations/institutions that had the space to house tens of sequencing equipment. The development of next generation sequencing revolutionized the process. It is now feasible to sequence the entire exome of multiple individuals in about 10 days. However, this meant that a massive amount of data needed to be filtered to identify the relevant alteration. This is presently the rate-limiting step in providing a convincing association between a genetic alteration and a human disorder.
人类基因组计划的完成提供了一个参考序列，研究人员可以将其与个体患者的序列进行比较，以期确定致病突变。然而，这仍然需要候选基因测试，或者使用桑格测序对多个基因进行非常有限的筛选。随着高通量桑格测序的出现，有可能对数百名患者进行数百个基因的改变筛查。这个过程既耗时又限于少数几个有空间容纳数十台测序设备的地点/机构。下一代测序的发展彻底改变了这一过程。现在，在大约 10 天内对多个个体的整个外显子组进行测序是可行的。然而，这意味着需要过滤大量的数据以识别相关的改变。这是目前在提供遗传改变与人类疾病之间的关联方面的一个限制步骤。
