...
  • 主题
高级搜索  >
历史搜索 清空历史
首页> 外文期刊>Annals of Human Genetics >Targeted next-generation sequencing on hirschsprung disease: A pilot study exploits DNA pooling
【24h】

Targeted next-generation sequencing on hirschsprung disease: A pilot study exploits DNA pooling

机译：靶向针对hirssprung疾病的下一代测序：一项初步研究利用DNA合并

获取原文
获取原文并翻译 | 示例
           
页面导航
  • 摘要
  • 著录项
  • 相似文献
  • 相关主题

摘要

To adopt an efficient approach of identifying rare variants possibly related to Hirschsprung disease (HSCR), a pilot study was set up to evaluate the performance of a newly designed protocol for next generation targeted resquencing. In total, 20 Chinese HSCR patients and 20 Chinese sex-matched individuals with no HSCR were included, for which coding sequences (CDS) of 62 genes known to be in signaling pathways relevant to enteric nervous system development were selected for capture and sequencing. Blood DNAs from eight pools of five cases or controls were enriched by PCR-based RainDance technology (RDT) and then sequenced on a 454 FLX platform. As technical validation, five patients from case Pool-3 were also independently enriched by RDT, indexed with barcode and sequenced with sufficient coverage. Assessment for CDS single nucleotide variants showed DNA pooling performed well (specificity/sensitivity at 98.4%/83.7%) at the common variant level; but relatively worse (specificity/sensitivity at 65.5%/61.3%) at the rare variant level. Further Sanger sequencing only validated five out of 12 rare damaging variants likely involved in HSCR. Hence more improvement at variant detection and sequencing technology is needed to realize the potential of DNA pooling for large-scale resequencing projects.
机译：为了采用一种有效的方法来鉴定可能与Hirschsprung病（HSCR）相关的稀有变体，我们进行了一项初步研究，以评估针对下一代靶向定性的新设计方案的性能。总共包括20例中国HSCR患者和20例没有HSCR的中国性匹配个体，为此选择了62个基因的编码序列（CDS），这些基因已知与肠道神经系统发育相关的信号传导途径用于捕获和测序。通过基于PCR的RainDance技术（RDT）富集了来自五个病例或对照的八个库的血液DNA，然后在454 FLX平台上进行了测序。作为技术验证，也通过RDT独立富集了病例Pool-3中的五名患者，用条形码索引并以足够的覆盖范围进行了测序。对CDS单核苷酸变体的评估表明，在常见变体水平上，DNA合并表现良好（特异性/敏感性为98.4％/ 83.7％）。但在稀有变异水平上相对较差（特异性/敏感性为65.5％/ 61.3％）。进一步的Sanger测序仅验证了HSCR可能涉及的12种罕见破坏性变体中的5种。因此，需要进一步改进变异检测和测序技术，以实现用于大规模重测序项目的DNA合并潜力。

著录项

相似文献

  • 外文文献
  • 中文文献
  • 专利
获取原文

客服邮箱：kefu@zhangqiaokeyan.com

京公网安备：11010802029741号 ICP备案号：京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有

  • 客服微信

  • 服务号