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Updated: May 25, 2026
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Targeted Next-generation Sequencing and Bioinformatics Pipeline to Evaluate Genetic Determinants of Constitutional Disease
Published on: April 4, 2018
Analysis of DNA sequence variants detected by high-throughput sequencing.
David R Adams1, Murat Sincan, Karin Fuentes Fajardo
1NIH Undiagnosed Diseases Program, NIH, Bethesda, Maryland, USA. david.adams@nih.gov
Human Mutation
|February 1, 2012
Summary
High-throughput sequencing (HTS) generates many DNA variants for diagnosing rare diseases. This study provides a practical roadmap for analyzing these variants, aiding researchers and clinicians in genetic diagnosis.
Area of Science:
- Genetics
- Bioinformatics
- Genomic Medicine
Background:
- High-throughput sequencing (HTS) is crucial for diagnosing rare and novel diseases.
- Analyzing the vast number of DNA variants from HTS requires robust filtering methods.
- A standardized analytic roadmap for HTS variant analysis is needed.
Purpose of the Study:
- To present a comprehensive analytic roadmap for high-throughput sequencing variant analysis.
- To share practical experiences and methods from the Undiagnosed Diseases Program.
- To empower researchers and clinicians to perform direct HTS data analysis.
Main Methods:
- Utilized high-throughput sequencing (HTS) exome data from over 30 families.
- Augmented exome data with high-density SNP-array data for each family.
- Detailed the theoretical and practical application of each analytic step for variant filtering.
Main Results:
- Successfully used HTS and SNP-array data to aid in clinical diagnoses for multiple families.
- Illustrated the analytic approach with example data for clarity.
- Developed a systematic process for filtering HTS-generated DNA sequence variants.
Conclusions:
- The presented analytic roadmap facilitates the interpretation of HTS data for rare disease diagnosis.
- This approach enables researchers and clinical geneticists to analyze HTS data effectively.
- Provides a valuable resource for advancing genetic diagnostics and understanding novel diseases.
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