摘要目的 总结功能、蛋白、基因诊断技术在慢性肉芽肿病(CGD)精准诊断中的价值.方法 回顾性分析2008年1月至2015年12月复旦大学附属儿科医院基因诊断明确的138例CGD患儿临床及实验室检查资料,总结流式细胞仪-二氢若丹明分析(DHR分析)检测的呼吸爆发功能刺激指数(SI),检测gp91蛋白表达情况,5个致病基因(CYBB、CYBA、NCF1、NCF2、NCF4)基因检测结果,并分析刺激指数、gp91蛋白、基因检测结果之间的相互关系.结果 138例基因诊断明确的CGD患儿中CYBB基因突变123例(89.1％)、CYBA基因突变4例(2.9％)、NCF1基因突变5例(3.6％)、NCF2基因突变6例(4.4％)、NCF4基因突变0例.SI范围为:0.8 ～60.5,第25、50、75百分位分别为1.7、2.7、4.7.112例CGD患儿检测gp91表达结果,其中100例不表达gp91,2例部分表达gp91,10例正常表达gp91.发现的未曾报道的突变位点包括:CYBB基因的c.76-77delTT、c.343-344delCA、c.481A ＞T、c.1152G＞C、c.1613G＞A,NCF2基因的c.137T＞G.CYBB突变CGD患儿中,gp91正常表达患儿SI高于gp91不表达者(14.6比2.5,t=44.21,P=0.004),NCF1基因突变者SI高于CYBB基因突变者(17.7比2.5,t=60.8,P=0.003).结论 流式细胞仪-DHR分析、gp91蛋白的流式检测方法为CGD的重要诊断方法,可指导进一步的基因检测.不同的基因突变及突变类型可影响呼吸爆发功能及gp91的蛋白水平.使用从功能、蛋白到基因水平的诊断技术可以精准有效诊断CGD.

abstractsObjective To evaluate the methods of flow cytometric-dihydrorhodamine 123 (DHR) analysis,gp91 protein detection,gene mutation analysis for the precise diagnosis of chronic granulomatous disease (CGD).Method Clinical and laboratory data of patients with CGD confirmed by gene mutation analysis from 2008 to 2015 in Children's Hospital of Fudan University were retrospectively reviewed.The results of respiratory burst,gp91 protein level,and gene mutations were analyzed.The relationships among these three methods were explored.Result A total of 138 patients of CGD with confirmed gene mutation were included in this study,of them,123 cases(89.1％) had CYBB gene mutation,4 cases(2.9％) had CYBA mutation,5 cases(3.6％) had NCF1 mutation and 6 cases(4.4％) had NCF2 mutation.The range of stimulatory index (SI) was 0.8-60.5,the 25 th,50 th,75th percent was 1.7,2.7,4.7;112 cases had the results of gp91,of them,100 with gp910,2 with gp91-,and 10 with gp91 + Six mutations,which were not reported before,were c.76-77delTF,c.343-344delCA,c.481A ＞T,c.1152G ＞ C,c.1613G ＞ A for CYBB gene,and c.137T ＞ G for NCF2 gene.Among CGD patients with CYBB mutation,SI of patients with gp91 + was higher than patients with gp910 14.6 vs.2.5 (t =44.21,P =0.004).Patients of NCF1 mutation had higher SI than patients with CYBB mutation,17.7 vs.2.5 (t =60.8,P =0.003).Conclusion Flow cytometric-DHR analysis and gp91 protein detection are important diagnostic methods for CGD,they could help the precise diagnosis of CGD.Different mutation types,different mutation genes could have impact on the results of respiratory burst and gp91 level.The application of diagnostic technology from function,protein to gene analysis could help precise diagnosis of CGD.