Clinical Validation of a Haplotyping Method with Next-Generation Sequencing

[摘要] To the Editor:One of the most persistent problems in clinical genetic testing is interpretation of compound heterozygotes. Particularly problematic are recessive genes for which compound heterozygosity is relatively common, such as CYP21A2 (cytochrome P450, family 21, subfamily A, polypeptide 2), the gene responsible for >90% of congenital adrenal hyperplasia (CAH)1 cases. Next-generation sequencing (NGS) methods provide a way to both sequence and phase genes; however, the techniques are largely qualitative and do not provide a quantitative measure of confidence in the cis/trans call. To date, an intuitive and reliable statistic to summarize the quality of a phase call is not available for diagnostic sequencing. Without such an indicator, incorrect phase calls can be made easily when sequence coverage is low or when errors are introduced during library preparation or amplification.Previously, we described a statistical method to generate probability scores and associated confidence intervals for each base in a tandem sequence of heterozygous positions (1). That method is based on stepwise analysis of sequential pairs of heterozygous loci. Although the method is functional, it suffers in regions where coverage is low and …

[学科分类] 过敏症与临床免疫学