RESUMO
The purpose of this paper is to describe how the authors used Internet services to generate a sampling frame for their on-going pilot study, which evaluates the psychometric properties of Chen's Nursing Home Tool, a community needs assessment instrument for nursing homes. The proposed sampling frame may be used to survey nursing home residents and family members, nursing home providers and community leaders for any selected community in the USA. The availability of a user-friendly sampling frame can assist health-care professionals to determine required and desired services and further contribute to the development of a more comprehensive health-service system.
Assuntos
Pesquisas sobre Atenção à Saúde , Internet , Avaliação das Necessidades , Casas de Saúde , Serviços de Saúde Comunitária , Humanos , Psicometria , Sistema de Registros , Estudos de AmostragemRESUMO
AIM: To develop a DNA-based diagnostic test for adrenoleukodystrophy (ALD) in a large New Zealand family. METHODS: Mutation screening of the X chromosome-linked ALD gene was undertaken by direct sequencing of PCR amplified products encompassing defined exons of the ALD gene. The identification of a mutation led to the development of a simple restriction enzyme digestion protocol of a PCR amplified product to identify those individuals with the mutation. RESULTS: A nonsense mutation, resulting in deduced premature termination of translation of the ALD gene product, was detected in exon 4 of the ALD gene in an affected male. This mutation was found in three obligate gene carriers in the same ALD family. A DNA-based test was established to identify this mutation by Bgl II digestion of a PCR amplified product encompassing exons 3 and 4 of the ALD gene. The DNA-based test was applied to a chorionic villus sampling for prenatal diagnosis. CONCLUSIONS: A simple DNA-based test has been developed for ALD in a large New Zealand family. This test provides a rapid means of determining carrier status and for undertaking prenatal diagnosis for ALD in this family.
Assuntos
Adrenoleucodistrofia/genética , DNA/genética , Adolescente , Adrenoleucodistrofia/diagnóstico , Adulto , Criança , Pré-Escolar , Amostra da Vilosidade Coriônica , Códon sem Sentido/genética , DNA/análise , Análise Mutacional de DNA , Éxons/genética , Feminino , Amplificação de Genes , Ligação Genética/genética , Testes Genéticos , Heterozigoto , Humanos , Masculino , Mutação/genética , Nova Zelândia , Reação em Cadeia da Polimerase , Biossíntese de Proteínas/genética , Transcrição Gênica/genética , Cromossomo X/genéticaRESUMO
Microsatellites of the dystrophin gene have been used extensively in the genetic analysis of Duchenne and Becker muscular dystrophy families. The microsatellites that have been reported to date are clustered within disparate regions of the dystrophin gene, specifically at the 5'-end and in the central rod-domain. YACs encompassing the gene were screened for further microsatellites to improve the density of available genetic markers. Four microsatellites were localized to defined regions of the dystrophin gene by the analysis of patient DNA samples, somatic cell hybrids and YACs. In addition, varying combinations of microsatellite loci were amplified in multiplex PCRs, which complement those loci that have been studied to date.
Assuntos
DNA Satélite/genética , Distrofina/genética , Reação em Cadeia da Polimerase , Sequência de Bases , Southern Blotting , Cromossomos Artificiais de Levedura , DNA/análise , DNA/genética , Primers do DNA , Humanos , Células Híbridas , Dados de Sequência Molecular , Valores de ReferênciaRESUMO
The Duchenne muscular dystrophy locus is remarkable in that it shows a high mutation rate and the majority of mutations found are deletions. These deletions are generated as meiotic as well as mitotic events and occur preferentially in the central region of the gene. Nothing is known so far about the mechanisms involved. This paper reports the first sequencing of deletion junctions in the dystrophin gene. The data from a study of two patients with deletions in the central region of dystrophin show the breakpoints to lie in regions of introns in which stretches of dA-dT are seen. The relationship between these observations and possible mechanisms for the mutations is discussed.
Assuntos
Deleção Cromossômica , Distrofina/genética , Distrofias Musculares/genética , Sequência de Bases , Southern Blotting , Clonagem Molecular , DNA , Análise Mutacional de DNA , Feminino , Humanos , Masculino , Dados de Sequência Molecular , Reação em Cadeia da Polimerase , Mapeamento por RestriçãoRESUMO
A cDNA clone coding for a mite allergen of mol wt approximately 14,000 has been isolated and its DNA sequence determined. The native component from mite extracts encoded by this DNA was identified by immunoprobing blots of mite body extract with human IgE eluted from the electroblotted cloned fusion polypeptides derived from the expressed cDNA clone. The clone encodes a polypeptide with a deduced mol wt of 17,460. The deduced amino acid sequence was not homologous to any known protein sequences and it contains no cysteine or tryptophan. On blots, 21 of 38 sera from mite-allergic subjects recognized the cloned material, and this recognition strongly correlated with IgE-binding to the native component on protein blots of mite extract.
