RESUMO
OBJECTIVE: To assess the practical value of BIOMED-2 primers in the diagnosis of ocular adnexal lymphoma by PCR. METHODS: DNA was extracted from 63 formalin-fixed paraffin-embedded (FFPE) ocular adnexal lymphoma specimens. The DNA quality was evaluated by PCR-based amplification of housekeeping gene beta-actin. IgH(B) and IgK(B) primers of BIOMED-2 standardized clonality analysis system were used to evaluate the immunoglobin gene rearrangements. PCR products were analyzed using capillary electrophoresis and GeneScan software. RESULTS: 76.2% (48/63) of FFPE samples produced amplifiable DNA for detection of Ig gene rearrangements.Positive detection rates by BIOMED-2 IgH(B) and IgK(B) primers were 79.2% (38/48) and 68.8% (33/48), respectively, with a combined positive detection rate of 91.7% (44/48). CONCLUSIONS: IgH(B) and IgK(B) primers of BIOMED-2 are suitable for the detection of clonal rearrangements of Ig gene using FFPE specimens of ocular adnexal lymphomas.
Assuntos
Actinas/genética , Neoplasias Oculares/genética , Rearranjo Gênico de Cadeia Pesada de Linfócito B , Rearranjo Gênico de Cadeia Leve de Linfócito B , Linfoma de Células B/genética , Adulto , Idoso , Primers do DNA , Neoplasias Oculares/diagnóstico , Neoplasias Oculares/patologia , Feminino , Humanos , Linfoma de Células B/diagnóstico , Linfoma de Células B/patologia , Linfoma Folicular/diagnóstico , Linfoma Folicular/genética , Linfoma Folicular/patologia , Masculino , Pessoa de Meia-Idade , Inclusão em Parafina , Projetos Piloto , Adulto JovemRESUMO
OBJECTIVE: To identify FBN1 gene mutations in a Chinese family with Marfan syndrome. METHODS: Four affected and two unaffected individuals in the family were recruited after informed consent. Five ml blood samples were drawn from each family member and genomic DNA was extracted. Mutations were detected by directly sequencing to the whole coding region and exon-intron boundaries of FBN1 gene. Polyphen program was used to predict the functional and structural changes of the mutant protein. RESULTS: We found all four affected individuals carried FBN1gene mutations, c.2261A > G (p.Y754C), in exon18 by sequence analysis, while two unaffected family members and 100 normal controls did not have this mutation. A PSIC score of 2.6 was acquired by Polyphen program analysis. CONCLUSION: Our study supports that FBN1 gene mutation, c.2261A > G (p.Y754C), is the underlying molecular pathogenesis of this family with Marfan syndrome. This mutation is identified for the first time in Chinese population.