Mutaitonal Analysis Of GBA In Gaucher Disease

BackgroundGaucher disease（GD） is an antosomal recessive lysosomal storage disease.Because of GBA gene mutation, glucocerebrosidase activity is low or deficient,making glucosylceramide store in monocyte-macrophages. The main manifestationsare hepatosplenomegaly、anemia、thrombocytopenia、fracture、growth retardation、repeating epilepsy and cerebellar ataxia. Gaucher disease has a poor prognosis,bringing the patients and their families big sorrow and increasing a big burdon on thesociety. So the conclusion of the clinical manifestations and bone marrow punctureresults for the diseases is particularly important for the early diagnosis. At the sametime, the genetic diagnose and genetic counseling are both more important for thereducing the occurrenct of Gaucher disease.ObjectiveIn this study, we summed up clinical manifestations and bone marrow punctureresults of two Gaucher disease children from two different families in our hospitalfrom2010and speculated the relationship between clinical manifestations and geneticmutations.MethodsAfter the consent of the two families, we collected the blood of the twoGaucher disease patients and their parents. At the same time, we collected the bloodof ten healthy children without blood lineage. After getting genomic DNA fromperipheral blood, we separately designed primers for the eleven exons and theirflanking sepuence of GBA. Using the forword and reverse primers, we sequenced the PCR products. At last, we aligned the sequencing results with the genomic DNAsequence of Genebank.Results1．In lineage1and10healthy children, the PCR products were consistent withthe expected fragment length and can be purified and sequenced.2．In lineage2，the product of PCR of exon9was shorter than the expectedfragment length, other PCR products and the PCR products of her parents wereconsistent with the expected fragment length and can be purified and sequenced.3． There were four mutations found in exon6of GBA in proband of case1andcase2, N188S （c.680A＞G） heterozygous mutation, N188K （c.681T＞G）heterozygous mutation, V191G（c.689T＞G）heterozygous mutation, S196P（c.703T＞C）heterozygous mutation，they were also found in their parents and10healthychildren.4．Deletion mutation c.1263₁317del55and D409H（c.1342G＞C）homozygousmutations were found in exon9of GBA in proband of case2，but not found in hisparents.Conclusion1．Mutations of GBA were detected in lineage1and2，these mutations werereported.2． N188S （c.680A＞G） heterozygous mutation, N188K （c.681T＞G）heterozygous mutation, V191G（c.689T＞G）heterozygous mutation, S196P（c.703T＞C）heterozygous mutation were found in proband1and2. It is reported that theyare in relation to pseudogene GBA. In order to identify them, we should redesignprimers of GBA exon6and undergo PCR amplifications.3．Deletion mutation c.1263₁317del55and D409H（c.1342G＞C）homozygousmutations found in lineage2were only found in proband of lineage2，consideringmutation found in lineage2were only found in proband of lineage2，consideringthese mutations were not from her parents but by herself. 4. This study revealed molecular genetic basis some Gaucher disease patients inmolecular level and enriched molecular genetic study of Gaucher disease in China,providing a feasible method for molecular genetic study of Gaucher disease.