GENE - Núcleo de Genética Médica, Belo Horizonte, Minas Gerais, Brazil.
PLoS One. 2013 Apr 19;8(4):e61328. doi: 10.1371/journal.pone.0061328. Print 2013.
Because of economic limitations, the cost-effective diagnosis of patients affected with rare microdeletion or microduplication syndromes is a challenge in developing countries. Here we report a sensitive, rapid, and affordable detection method that we have called Microdeletion/Microduplication Quantitative Fluorescent PCR (MQF-PCR). Our procedure is based on the finding of genomic regions with high homology to segments of the critical microdeletion/microduplication region. PCR amplification of both using the same primer pair, establishes competitive kinetics and relative quantification of amplicons, as happens in microsatellite-based Quantitative Fluorescence PCR. We used patients with two common microdeletion syndromes, the Williams-Beuren syndrome (7q11.23 microdeletion) and the 22q11.2 microdeletion syndromes and discovered that MQF-PCR could detect both with 100% sensitivity and 100% specificity. Additionally, we demonstrated that the same principle could be reliably used for detection of microduplication syndromes, by using patients with the Lubs (MECP2 duplication) syndrome and the 17q11.2 microduplication involving the NF1 gene. We propose that MQF-PCR is a useful procedure for laboratory confirmation of the clinical diagnosis of microdeletion/microduplication syndromes, ideally suited for use in developing countries, but having general applicability as well.
由于经济限制，在发展中国家，对于患有罕见微缺失或微重复综合征的患者，经济有效的诊断是一个挑战。在这里，我们报告了一种敏感、快速且经济实惠的检测方法，我们称之为微缺失/微重复定量荧光 PCR（MQF-PCR）。我们的方法基于发现与关键微缺失/微重复区域的片段具有高度同源性的基因组区域。使用相同的引物对对这两个区域进行 PCR 扩增，建立了扩增子的竞争动力学和相对定量，就像基于微卫星的定量荧光 PCR 一样。我们使用了两种常见的微缺失综合征（Williams-Beuren 综合征（7q11.23 微缺失）和 22q11.2 微缺失综合征）的患者，并发现 MQF-PCR 可以 100%灵敏和 100%特异地检测到这两种综合征。此外，我们通过使用 Lubs（MECP2 重复）综合征和涉及 NF1 基因的 17q11.2 微重复的患者，证明了相同的原理可以可靠地用于微重复综合征的检测。我们提出 MQF-PCR 是一种用于实验室确认微缺失/微重复综合征临床诊断的有用方法，非常适合发展中国家使用，但也具有普遍适用性。
