RESUMO
A simple method for detecting pathogenic microorganisms in clinical samples has been developed. This method is based on identification of specific nucleotide sequences by Pt[(dien)Cl] Cl-labelled DNA probes and simultaneous immunochemical control of total DNA content in each clinical sample. Such control is provided by a simple semiquantitative enzyme-linked immunoassay using antibodies to denatured DNA; its implementation requires the same procedures, materials and solutions used in the hybridization analysis. The information about the DNA content in the sample is necessary for adequate interpretation of results of hybridization analysis. Pt[(dien)Cl]Cl for the probe and affinity antibodies to DNA-Pt[(dien)Cl] Cl and rabbit immunoglobulin antibodies conjugated to phosphatase for DNA hybrid detection were used. The results of hybridization analysis show a good correlation with those of the PCR-test. The method is simple, relatively inexpensive and fit for population monitoring.
Assuntos
Chlamydia trachomatis/isolamento & purificação , Cisplatino/análogos & derivados , Sondas de DNA , Ureaplasma urealyticum/isolamento & purificação , Infecções por Chlamydia/diagnóstico , Chlamydia trachomatis/genética , Cisplatino/química , DNA Bacteriano/isolamento & purificação , Feminino , Humanos , Masculino , Hibridização de Ácido Nucleico , Reação em Cadeia da Polimerase , Infecções por Ureaplasma/diagnóstico , Ureaplasma urealyticum/genéticaRESUMO
Patients with Duchenne muscular dystrophy were analyzed using the method of polymerase chain reaction in order to reveal deletions in the dystrophin gene. Deletions of different lengths and locations were detected in 28 of 78 ill boys. The highest number of deletions was detected in the 3'-end of the gene (the 45-50th exons).
Assuntos
Deleção Cromossômica , Distrofias Musculares/genética , Adolescente , Criança , Pré-Escolar , Primers do DNA , Distrofina/genética , Eletroforese em Gel de Poliacrilamida , Éxons/genética , Humanos , Masculino , Dados de Sequência Molecular , Distrofias Musculares/diagnóstico , Reação em Cadeia da Polimerase/métodosRESUMO
Multiplex polymerase chain reaction was carried out with the material from 68 patients suffering from Duchenne muscular dystrophy in Moscow and Leningrad clinics. Six pairs of oligoprimers were used. Deletions were detected in the material from 22 patients. A new type of deletion was found. Data on deletion frequencies and spectrum were compared with the results published by other authors.
Assuntos
Distrofina/genética , Deleção de Genes , Distrofias Musculares/genética , Criança , Mapeamento Cromossômico , Eletroforese em Gel de Ágar , Amplificação de Genes , Humanos , Masculino , Reação em Cadeia da PolimeraseRESUMO
Routinely, we detect 0,1 pg of plasmid DNA using the nonradioactive DNA labeling and detection kit produced by Boehringer Mannheim (FRG). Using the kit we have determined the carrier status of a woman in a family with a case of Duchenne muscular dystrophy by the blot hybridization technique.
Assuntos
Sondas de DNA , Distrofias Musculares/diagnóstico , Southern Blotting , Feminino , Triagem de Portadores Genéticos , Humanos , Linhagem , PlasmídeosRESUMO
Two prenatal diagnoses were carried out by the technique of intragenic polymorphous marker detecting heterozygosity in pregnant women in the families with cases of Duchenne muscular dystrophy. In both cases the DNA fragment from pERT87-15 region was amplified. This fragment includes a polymorphous site in BamHI region of recognition. DNA analyses of the families members have been made and the genetical risk has been calculated by the Bayes method. The prognoses for both fetuses are good.
Assuntos
Distrofias Musculares/diagnóstico , Diagnóstico Pré-Natal , Sequência de Bases , Desoxirribonuclease BamHI , Feminino , Doenças Fetais/diagnóstico , Amplificação de Genes , Marcadores Genéticos , Humanos , Dados de Sequência Molecular , Linhagem , Polimorfismo de Fragmento de Restrição , GravidezRESUMO
Duchenne muscular dystrophy carrier detection has been performed by using probes XJ1.1 (intragenic probe) and probe 754 for a girl. The carrier probability was estimated by means of a computer program GenRisk combining pedigree and DNA-probe data and turned out to be 95%.
Assuntos
Sondas de DNA , Heterozigoto , Distrofias Musculares/genética , Alelos , Feminino , Humanos , Masculino , Hibridização de Ácido Nucleico , Linhagem , SoftwareRESUMO
Seven families with histories of Duchenne's muscular dystrophy underwent DNA diagnosis. The daughters of those consulted were examined for the carriage in 4 families. Their carriage was rejected or confirmed. Prenatal diagnosis was made in 2 families. In another family an abortion preceded obtaining molecular-genetic evidence. Probes 754, p20, XJI.I and primers for amplification of the site pERI87-15 containing a polymorphic locus were employed. The genetic risk was assessed using the computer program GenRisk adjusted for family history and DNA test allowances.
Assuntos
Sondas de DNA , Triagem de Portadores Genéticos/métodos , Distrofias Musculares/genética , Diagnóstico Pré-Natal/métodos , Adulto , Alelos , Criança , Feminino , Humanos , Masculino , Distrofias Musculares/diagnóstico , Linhagem , Gravidez , Cromossomo X/ultraestruturaRESUMO
A fraction of synaptonemal complexes (SC) isolated from mouse spermatocytes has been electrophoretically purified in agarose gel. The DNA from the SC fraction constitutes approximately 0.5% of total nuclear DNA, and its molecules have length heterogeneity from 1 k.b. to 20 k.b. The content of beta-globin gene is the same in DNA from the SC fraction and in total nuclear DNA. The specificity of DNA from the SC fraction is manifested by higher contents of the repeated alternative sequences GT/CA and B1-sequence that is probably due to the processes of genetic meiotic recombination.
