Identification of Deletion Carriers in X-Linked Chronic Granulomatous Disease by Real-Time PCR
Identification of Deletion Carriers in X-Linked Chronic Granulomatous Disease by Real-Time PCR
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DOI：
10.1089/gtmb.2009.0074
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发表时间：
2009-12-01
影响因子：
1.4
通讯作者：
Finocchi, Andrea
中科院分区：
文献类型：
--
作者：
Chiriaco, Maria;Di Matteo, Gigliola;Finocchi, Andrea
慢性肉芽肿病（CGD）是一种罕见的影响先天免疫系统的原发性免疫缺陷。即使功能测试可以诊断先天性先天性遗传病，但分子缺陷的识别对于向有患先天性先天性遗传病风险的家庭成员提供咨询和产前诊断至关重要。x连锁型出现在65%的CGD患者中。这是由于CYBB基因的突变，其中高达12%是由大量缺失引起的。CGD携带者通常是健康的，分子分析是揭示其携带者状态的必要手段。本研究的目的是应用基因剂量法，使用SYBR绿色定量实时聚合酶链反应（RT-PCR），量化CYBB基因区域严重缺失的携带者和非携带者的基因组拷贝数。我们研究了两种不同的CYBB基因扩增产物的表达，结果证实该基因在载体样品中的表达高度降低。结果经连锁分析和荧光原位杂交证实。实时定量PCR快速简便，我们建议将其作为一种新的常规诊断方法，用于检测CYBB基因区域缺失的CGD携带者。
Chronic granulomatous disease (CGD) is a rare primary immunodeficiency affecting the innate immune system. Even if functional tests address the diagnosis of CGD, the identification of a molecular defect is essential for counselling family members at risk for being CGD carriers and for prenatal diagnosis. The X-linked form occurs in 65% of CGD patients. It is due to mutations in the CYBB gene, up to 12% of which are caused by large deletions. CGD carriers are usually healthy, and molecular analysis is essential to reveal their carrier status. The aim of this study was to apply a gene dosage approach, using SYBR green quantitative real-time polymerase chain reaction (RT-PCR), to quantify the genomic copy number in carriers and noncarriers of gross deletions covering the region of the CYBB gene. We studied the expression of two different amplification products of the CYBB gene, and the results confirmed a highly reduced expression of the gene in the carrier samples. The results were confirmed by linkage analysis and fluorescence in situ hybridization. Quantitative real-time PCR is fast and simple to perform, and we propose it as a new routine diagnostic approach to detect CGD carriers of deletions covering the region spanning the CYBB gene.