Assuntos
Neoplasias da Mama/metabolismo , Neoplasias da Mama/patologia , Imuno-Histoquímica/métodos , Receptor ErbB-2/análise , Receptores de Estrogênio/análise , Receptores de Progesterona/análise , Análise Serial de Tecidos/métodos , Adulto , Idoso , Idoso de 80 Anos ou mais , Feminino , Humanos , Pessoa de Meia-Idade , Valor Preditivo dos Testes , Receptor ErbB-2/metabolismo , Receptores de Estrogênio/metabolismo , Receptores de Progesterona/metabolismo , Reprodutibilidade dos Testes , SudãoRESUMO
AIMS AND BACKGROUND: Copy number variations (CNVs) contribute to genome variability and their pathogenic role is becoming evident in an increasing number of human disorders. Commercial assays for routine diagnosis of CNVs are available only for a fraction of known genomic rearrangements. Thus, it is important to develop flexible and cost-effective methods that can be adapted to the detection of CNVs of interest, both in research and clinical settings. METHODS: We describe a new multiplex PCR-based method for CNV analysis that exploits automated microfluidic capillary electrophoresis through lab-on-a-chip technology (LOC-CNV). We tested the reproducibility of the method and compared the results obtained by LOC-CNV with those obtained using previously validated semiquantitative assays such as multiplex ligation-dependent probe amplification (MLPA) and nonfluorescent multiplex PCR coupled to HPLC (NFMP-HPLC). RESULTS: The results obtained by LOC-CNV in control individuals and carriers of pathogenic MLH1 or BRCA1 genomic rearrangements (losses or gains) were concordant with those obtained by previously validated methods, indicating that LOC-CNV is a reliable method for the detection of genomic rearrangements. CONCLUSION: Because of its advantages with respect to time, costs, easy adaptation of previously developed multiplex assays and flexibility in novel assay design, LOC-CNV may represent a practical option to evaluate relative copy number changes in genomic targets of interest, including those identified in genome-wide analyses.
Assuntos
Variações do Número de Cópias de DNA , Dispositivos Lab-On-A-Chip , Reação em Cadeia da Polimerase Multiplex , Proteínas Adaptadoras de Transdução de Sinal/genética , Proteína BRCA1/genética , Neoplasias da Mama/genética , Cromatografia Líquida de Alta Pressão , Neoplasias Colorretais Hereditárias sem Polipose/genética , Europa (Continente) , Rearranjo Gênico , Heterozigoto , Humanos , Dispositivos Lab-On-A-Chip/economia , Dispositivos Lab-On-A-Chip/estatística & dados numéricos , Reação em Cadeia da Polimerase Multiplex/métodos , Proteína 1 Homóloga a MutL , Proteínas Nucleares/genética , Reprodutibilidade dos TestesRESUMO
The African Society of Human Genetics (AfSHG), founded in 2003 with its inaugural meeting in Accra, Ghana,1 has the stated missions of (1) disseminating information about human genetics research in Africa, (2) establishing a mentorship network providing educational resources, including the development of appropriate technology transfer, (3) providing advocacy for human genetic research in Africa, and (4) encouraging collaborative research. Despite its young age, the AfSHG has developed a strong cadre of active researchers, both within and outside of Africa, with more than 400 members (from 16 countries across Africa as well as 8 other countries), and has held six successful meetings, five in Africa and one in the United States.