高通量测序在临床分子诊断中的应用与展望

doi:10.3969/j.issn.1673-8640.2017.04.002

摘要/Abstract

摘要：

高通量测序又称下一代测序（NGS）,是一种新型的遗传学筛查和诊断技术,它的不断革新加速了人们对遗传学标志物及疾病分子机制的认识,特别是针对复杂遗传性疾病。NGS技术的出现促使临床基因诊断逐渐从单基因时代跨越到多基因时代。随着靶向基因组测序、全外显子组测序以及全基因组测序项目在临床中的广泛应用,医务工作者将逐渐改变对复杂遗传性疾病的管理策略,最终将依据遗传学检测结果对每一位患者进行2次分类并给予精准治疗。另外,生物信息学分析方法的不断更新和便捷的数据分析软件包也使NGS在临床诊断中的应用更加广泛。文章中主要论述了现有NGS的主流平台、现阶段NGS在临床分子诊断中的主要应用策略以及应用NGS检测序列变异的数据分析方法。

关键词: 高通量测序, 复杂遗传性疾病, 临床基因诊断, 生物信息学

Abstract:

：High-throughput sequencing,next generation sequencing（NGS）,is a new type of genetic screening technology. Its innovation accelerates the understanding of genetic markers and molecular mechanism,and also promotes clinical gene diagnosis from single gene to multi-genes,especially for complex genetic diseases. As the new sequencing technology used in clinic,doctors will change the management of genetic diseases,and patients will be placed into different groups eventually based on genetic diagnosis. In addition,with the improvement of bioinformatics and package,the application of NGS in clinic will be more routine. This review discusses the major platform and performance of NGS and its application in clinic and the process of NGS analysis of sequence variations.

Key words: High-throughput sequencing, Complex genetic diseases, Clinical gene diagnosis, Bioinformatics

中图分类号:

R393

姜晓峰. 高通量测序在临床分子诊断中的应用与展望[J]. 检验医学, 2017, 32(4): 250-254.

JIANG Xiaofeng. Application and prospect of high-throughput sequencing in clinical molecular diagnosis[J]. Laboratory Medicine, 2017, 32(4): 250-254.

图/表 2

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平台	扩增方式	测序原理	读段长度（bp）	运行时间（h）	费用/Gb（美元）	错误率（%）
454 GS Junior	乳滴PCR	焦磷酸测序	25~700	20	19 540	0.5
IlluminaMiSeq	桥式PCR	合成测序	300~600	21~55	109~996	0.2
Illumina HiSeq2500	桥式PCR	合成测序	125~250	60~144	30~90	0.2
IlluminaHiSeq X Ten	桥式PCR	合成测序	150	72	7	0.2
SOLiD 5500xl	乳滴PCR	链接测序	50~75	144	68	0.1
Ion PGM	乳滴PCR	半导体测序	200~400	2.3~7.3	460~788	1.0
Ion Proton	乳滴PCR	半导体测序	最大200	2~4	11.0~81.6	1.0
PacBio RS II	NONE	SMRT测序	8 500	4	1 111	15.0

平台	扩增方式	测序原理	读段长度（bp）	运行时间（h）	费用/Gb（美元）	错误率（%）
454 GS Junior	乳滴PCR	焦磷酸测序	25~700	20	19 540	0.5
IlluminaMiSeq	桥式PCR	合成测序	300~600	21~55	109~996	0.2
Illumina HiSeq2500	桥式PCR	合成测序	125~250	60~144	30~90	0.2
IlluminaHiSeq X Ten	桥式PCR	合成测序	150	72	7	0.2
SOLiD 5500xl	乳滴PCR	链接测序	50~75	144	68	0.1
Ion PGM	乳滴PCR	半导体测序	200~400	2.3~7.3	460~788	1.0
Ion Proton	乳滴PCR	半导体测序	最大200	2~4	11.0~81.6	1.0
PacBio RS II	NONE	SMRT测序	8 500	4	1 111	15.0

数据源	描述
RefSeq	由NCBI提供的具有生物学意义的非冗余序列数据库
ASAP II	剪切变异体数据库
ASPicDB	人类基因剪切模式数据库
ASTD	由可变剪切或可变起始或终止位点产生的可变转录物数据库
dbSNP	NCBI上的变异体目录
OMIM	人类遗传疾病及其致病基因数据库
SNPeffect	注释单核苷酸多态性影响的数据库
Swiss-prot	非冗余的蛋白质序列数据库
HGMD	人类基因突变数据库
COSMIC	癌症体细胞突变数据库

数据源	描述
RefSeq	由NCBI提供的具有生物学意义的非冗余序列数据库
ASAP II	剪切变异体数据库
ASPicDB	人类基因剪切模式数据库
ASTD	由可变剪切或可变起始或终止位点产生的可变转录物数据库
dbSNP	NCBI上的变异体目录
OMIM	人类遗传疾病及其致病基因数据库
SNPeffect	注释单核苷酸多态性影响的数据库
Swiss-prot	非冗余的蛋白质序列数据库
HGMD	人类基因突变数据库
COSMIC	癌症体细胞突变数据库