Laboratory diagnosis of chronic granulomatous disease

Abstract

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). 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. 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 gp91(0,) 2 with gp91(-), and 10 with gp91(+) . Six mutations, which were not reported before, were c. 76-77delTT, 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 gp91(0) 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). 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.