Bermudez Mercedes G, Piyamongkol Wirawit, Tomaz Susana, Dudman Evelyn, Sherlock Jon K, Wells Dagan
The Institute for Reproductive Medicine and Science, Saint Barnabas Medical Center, West Orange, NJ 07052, USA.
Prenat Diagn. 2003 Aug;23(8):669-77. doi: 10.1002/pd.658.
There is increasing interest in the use of preimplantation genetic diagnosis (PGD) as an alternative to routine prenatal diagnosis. However, the costs associated with development and testing of new PGD protocols have forced some PGD centres to limit the number of diseases for which PGD is offered. One of the main factors in the design of new protocols, which affects cost and accuracy, is the choice of the mutation-detection technique. We have assessed the reliability of DNA sequencing and mini-sequencing for clinical diagnosis at the single-cell level and have found them to be rapid and accurate. Extensive optimisation for individual mutations is not usually necessary when employing these versatile techniques and consequently a smaller investment of time and resources should be required during development of new protocols. Additionally, we report single-cell protocols for the diagnoses of cystic fibrosis, sickle cell anaemia and beta-thalassaemia, which utilise mini-sequencing. Unlike most mutation-detection techniques, mini-sequencing permits analysis of very small DNA fragments. Small amplicons experience low allele dropout (ADO) rates, and consequently this approach could potentially improve the reliability of PGD.
人们对使用植入前基因诊断（PGD）作为常规产前诊断的替代方法越来越感兴趣。然而，与新的PGD方案的开发和测试相关的成本迫使一些PGD中心限制提供PGD检测的疾病数量。新方案设计中的一个主要因素，它会影响成本和准确性，是突变检测技术的选择。我们评估了DNA测序和微测序在单细胞水平上用于临床诊断的可靠性，发现它们快速且准确。在使用这些通用技术时，通常不需要对个别突变进行广泛优化，因此在开发新方案时所需的时间和资源投入应该更少。此外，我们报告了利用微测序诊断囊性纤维化、镰状细胞贫血和β地中海贫血的单细胞方案。与大多数突变检测技术不同，微测序允许分析非常小的DNA片段。小扩增子的等位基因脱扣（ADO）率低，因此这种方法可能会提高PGD的可靠性。
