Assuntos
Feto , Diagnóstico Pré-Natal , Gravidez , Feminino , Humanos , Sequenciamento do Exoma , Hungria , Análise em MicrossériesRESUMO
Next generation sequencing (NGS) is a rapidly developing area in genetics. Utilizing this technology in the management of disorders with complex genetic background and not recurrent mutation hot spots can be extremely useful. In this study, we applied NGS, namely semiconductor sequencing to determine the most significant osteogenesis imperfecta-related genetic variants in the clinical practice. We selected genes coding collagen type I alpha-1 and-2 (COL1A1, COL1A2) which are responsible for more than 90% of all cases. CRTAP and LEPRE1/P3H1 genes involved in the background of the recessive forms with relatively high frequency (type VII and VIII) represent less than 10% of the disease. In our six patients (1-41 years), we identified 23 different variants. We found a total of 14 single nucleotide variants (SNV) in COL1A1 and COL1A2, 5 in CRTAP and 4 in LEPRE1. Two novel and two already well-established pathogenic SNVs have been identified. Among the newly recognized mutations, one results in an amino acid change and one of them is a stop codon. We have shown that a new full-scale cost-effective NGS method can be developed and utilized to supplement diagnostic process of osteogenesis imperfecta with molecular genetic data in clinical practice.
Assuntos
Sequenciamento de Nucleotídeos em Larga Escala , Osteogênese Imperfeita/genética , Adolescente , Adulto , Densidade Óssea , Criança , Pré-Escolar , Colágeno Tipo I/genética , Cadeia alfa 1 do Colágeno Tipo I , Proteínas da Matriz Extracelular/genética , Feminino , Humanos , Lactente , Masculino , Glicoproteínas de Membrana/genética , Chaperonas Moleculares , Osteogênese Imperfeita/patologia , Polimorfismo de Nucleotídeo Único , Prolil Hidroxilases , Proteoglicanas/genética , Análise de Sequência de DNARESUMO
INTRODUCTION: Hyperhomocysteinemia is an independent risk factor for cardiovascular morbidity. AIM: The study was designed to evaluate the total homocysteine level and MTHFR C677T polymorphism frequency of 122, healthy, young adults who had increased risk for cardiovascular disease. The serum levels of folic acid and vitamin B12 were also measured. METHODS: Immunoassay, PCR-RFLP methods were used. The statistical analysis was performed by SPSS program. RESULTS: The frequency of the gene-polymorphism was not different significantly in the study group compared to a Hungarian neonatal sample: although in the increased risk group the frequency of homozygous 677TT polymorphism was higher (14.8%), and heterozygosity was smaller (41%). There was no association between MTHFR gene polymorphism and homocysteine levels. A significant negative correlation was found between the folic acid and homocysteine, and between the vitamin B12 and homocysteine levels correlating with the literature. The mean serum total homocysteine level of the group without vitamin supplementation (n: 86) was 9.8 +/- 3.3 micromol/l, while in the other group with vitamin uptake (n: 36) this level was 7.5 +/- 3.0 micromol/l. There was a significant difference between the homocysteine levels of men and women. CONCLUSION: The results of the study correlate with the literature. It would be useful to call the attention of the Hungarian population to the importance of vitamin supply.
