Clinical Features And Molecular Anasysis Of Eight Patients With Chronic Granulomatous Disease In China

Objective To explore the clinical manifestations and the related genemutations of8patients with chronic granulomatous disease in Guangxi province,and the relationship between them.Methods Clinical data of8Chinese CGD patients are collected, who werediagnosed or treated at the first affiliated hospital of Guangxi MedicalUniversity during2007to2014. Analysis of blood routine, immunoglobulin test,lymphatic cellular sub-group, histopathology and radiology features wereperformed, Neutrophils respiratory burst assay was detected by Flow cytometryfor patients and their family members. Gene analysis was examined on6casesof them （P1has diagnosed at Children’s Hospital of Chongqing MedicalUniversity, P2has been reported in Yang Xiaoxu’s paper）: Total RNA andGenome DNA which was extracted from fresh whole blood, and amplificationCYBB and NCFI by reverse transcription polymerase chain reaction （RT-PCR）, PCR products were purified and bidirectional direct sequencing and thesequences were subsequently compared with reported CYBB and NCF1sequences in the GeneBank. If a special mutation was identified in CYBB orNCFI, the genomic DNA was amplified to confirm.Results （1） Clinical phenotype:8patients were all male, the average onsetage was50days old, the average diagnostic age was14months old.8cases allsuffered fever, all had recurrent pulmonary infection or severe pneumonia,6cases suffered skin abscesses and/or perianal abscesses,6cases sufferedlymphadenitis and/or hepatosplenomegaly, tuberculosis infection/（pulmonary）tuberculosis in4cases,2patients had growth and developmental retardation,hemophagocytic lymphollistiocytosis occurred in1case.3patients had positivefamily history. CGD patients all have elevated WBC,5patients withmild-moderate anemia; immuneglobulin and lymphatic cellular sub-group hadbeen test in6patients, lgA were all increased,5cases with IgG and lgMincreased; total T cells and lymphocytes CD4+T cells were generally normal,CD8+T cells were normal in2cases, and lower in4cases; NK cells werenormal in1case, increased in1case, and decreased in4cases. NAPDH oxidasefunction detected by flow cytometry were all significantly decreased.（2） Geneanalysis in6cases:1case was NCF1gene mutation（c.75₇6delGT）, it’s ahotspot mutation which has been reported; other5cases were CYYB genemutations,1missense mutation case （c.1235G＞T） had been reported before,the rest4cases were novel mutations had not been reported,1case was deletionmutation (c.1465₁468delATCA, and3cases were splicing mutation（c.1465₁468delATCA, g.IVS1-2A＞C, g.IVS71G＞T, g.IVS5-2A＞T）.Conclusion （1） children with chronic granulomatous disease had varied and severe clinical manifestations, routine laboratory tests lack of specificity todiagnosis, neutrophil respiratory burst test peripheral blood and gene analysiscan be used to rapid diagnosis for this disease.（2） In this study, there are4novelgene mutation in CYBB,1hotspot mutation in NCF1about CGD, there mightbe some relationship between the clinical phenotype and genotype, but thenumber of cases were too small to make a conclusion, it still need furtherresearch.