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目的:通过分析12例X连锁慢性肉芽肿病(X-CGD)的CYBB基因变异,探讨基因诊断在确诊该病中的价值。方法:根据患儿临床特征及四唑氮蓝试验(NBT)初步筛查结果:,运用流式细胞术中性粒细胞呼吸爆发试验,检测11例X-CGD患者及家系成员的中性粒细胞氧化功能,采用PCR及RT-PCR直接测序法,分析12例患儿及其家系外周血细胞的CYBB基因,并为1例CYBB突变基因携带者的胎儿分析羊水细胞的CYBB基因。结果:患儿均有反复感染史,NBT显著降低(0～6%),11例患儿中性粒细胞氧化指数(NOI)为1～2,明显低于正常人(>100),初步诊断为X-CGD;12例患儿中,11例存在CYBB基因突变:缺失突变4例,拼接错误突变4例,无义突变2例及错义突变1例。其中7种突变(1341delT,1182delC,1091delG,IVS10-2A>C,IVS10+2dupT,497-498ins115bp,591T>C)为新型突变。除了例3患儿母亲不是携带者,例9患儿母亲是否为携带者尚不明确,其余9个患儿母亲均为携带者,例6母亲的胎儿正常。结论:CYBB基因分析有助于X-CGD患儿的进一步明确诊断,在检测携带者及产前诊断中起着重要作用,并为干细胞移植治疗患者提供依据。
Objective: To investigate the value of gene diagnosis in diagnosing the disease by analyzing the CYBB gene mutation in 12 cases of X-linked chronic granulomatosis (X-CGD). Methods: According to the clinical features of children and the preliminary screening results of NBT, we used flow cytometry neutrophil respiratory burst test to detect the expression of neutrophils in 11 patients with X-CGD and their family members The CYBB gene of peripheral blood cells of 12 children and their families was analyzed by PCR and RT-PCR. The CYBB gene of amniotic fluid cells was analyzed by one fetus with CYBB mutation carrier. Results: There was a history of recurrent infections in all children, NBT was significantly reduced (0-6%), and 11 patients had a neutrophil oxidative index (NOI) of 1-2, which was significantly lower than that of normal subjects (> 100) Among the 12 patients, CYBB mutations were found in 11 cases: deletion mutation in 4 cases, misdirect mutation in 4 cases, nonsense mutation in 2 cases and missense mutation in 1 case. Seven of these mutations (1341delT, 1182delC, 1091delG, IVS10-2A> C, IVS10 + 2dupT, 497-498ins115bp, 591T> C) were novel mutations. Except for case 3, the mother of a child is not a carrier. In case 9, it is unclear whether the mother is a carrier or not, and the remaining 9 mothers are carriers. For example, the mother’s fetus is normal. Conclusion: CYBB gene analysis is helpful for the further diagnosis of X-CGD in children. It plays an important role in the detection of carriers and prenatal diagnosis, and provides the basis for the treatment of patients with stem cell transplantation.