Sanger sequencing is no longer always necessary based on a single-center validation of 1109 NGS variants in 825 clinical exomes.
Sanger sequencing is no longer always necessary based on a single-center validation of 1109 NGS variants in 825 clinical exomes.
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DOI：
10.1038/s41598-021-85182-w
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发表时间：
2021-03-11
影响因子：
4.6
通讯作者：
Ayuso C
中科院分区：
文献类型：
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作者：
Arteche-López A;Ávila-Fernández A;Romero R;Riveiro-Álvarez R;López-Martínez MA;Giménez-Pardo A;Vélez-Monsalve C;Gallego-Merlo J;García-Vara I;Almoguera B;Bustamante-Aragonés A;Blanco-Kelly F;Tahsin-Swafiri S;Rodríguez-Pinilla E;Minguez P;Lorda I;Trujillo-Tiebas MJ;Ayuso C
尽管下一代测序(NGS)的准确性有所提高，但人们普遍认为，在报告之前，需要使用桑格测序对变异进行验证。所有NGS变种的验证大大增加了临床诊断的周转时间和成本。我们在825个临床外显子的1109个变种中全面评估了这一需求，这是迄今为止使用Illumina化学方法评估的最大样本集。在100%的符合率下，我们得出结论，Sanger测序作为一种内部质量控制是非常有用的，但作为一种高质量的单核苷酸和小插入/缺失变体的验证方法还不够。在停止桑格确认研究之前，实验室可能会验证并建立自己的阈值。我们还扩展和验证了20个样本中外显子测序检测到的23个拷贝数变异，符合率为95.65%(22/23)。
Despite the improved accuracy of next-generation sequencing (NGS), it is widely accepted that variants need to be validated using Sanger sequencing before reporting. Validation of all NGS variants considerably increases the turnaround time and costs of clinical diagnosis. We comprehensively assessed this need in 1109 variants from 825 clinical exomes, the largest sample set to date assessed using Illumina chemistry reported. With a concordance of 100%, we conclude that Sanger sequencing can be very useful as an internal quality control, but not so much as a verification method for high-quality single-nucleotide and small insertion/deletions variants. Laboratories might validate and establish their own thresholds before discontinuing Sanger confirmation studies. We also expand and validate 23 copy number variations detected by exome sequencing in 20 samples, observing a concordance of 95.65% (22/23).
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影响因子：
9.3
作者：
Beck TF;Mullikin JC;NISC Comparative Sequencing Program;Biesecker LG
通讯作者：
Biesecker LG
影响因子：
2
作者：
Yao, Ruen;Yu, Tingting;Shen, Yiping
通讯作者：
Shen, Yiping
影响因子：
4.1
作者：
Kerkhof, Jennifer;Schenkel, Laila C.;Sadikovic, Bekim
通讯作者：
Sadikovic, Bekim
DOI：
10.1038/s41436-018-0295-y
发表时间：
2019-05
期刊：
Genetics in medicine : official journal of the American College of Medical Genetics
影响因子：
--
作者：
Gross AM;Ajay SS;Rajan V;Brown C;Bluske K;Burns NJ;Chawla A;Coffey AJ;Malhotra A;Scocchia A;Thorpe E;Dzidic N;Hovanes K;Sahoo T;Dolzhenko E;Lajoie B;Khouzam A;Chowdhury S;Belmont J;Roller E;Ivakhno S;Tanner S;McEachern J;Hambuch T;Eberle M;Hagelstrom RT;Bentley DR;Perry DL;Taft RJ
通讯作者：
Taft RJ
DOI：
10.1038/gim.2013.183
发表时间：
2014-07
期刊：
Genetics in medicine : official journal of the American College of Medical Genetics
影响因子：
--
作者：
通讯作者：
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