RESUMO
We used an infrared (IR) automated fluorescence monolaser sequencer for the analysis of 13 autosomal short tandem repeat (STR) systems (TPOX, D3S1358, FGA, CSF1PO, D5S818, D7S820, D8S1179, TH01, vWA, D13S317, D16S359, D18S51, D21S11) and the X-Y homologous gene amelogenin system. These two systems represent the core of the combined DNA index systems (CODIS). Four independent multiplex reactions, based on the polymerase chain reaction (PCR) technique and on the direct labeling of the forward primer of every primer pair, with a new molecule (IRDye800), were set up, permitting the exact characterization of the alleles by comparison with ladders of specific sequenced alleles. This is the first report of the whole analysis of the STRs of the CODIS core using an IR automated DNA sequencer. The protocol was used to solve paternity/maternity tests and for population studies. The electrophoretic system also proved useful for the correct typing of those loci differing in size by only 2 bp. A sensibility study demonstrated that the test can detect an average of 10 pg of undegraded human DNA. We also performed a preliminary study analyzing some forensic samples and mixed stains, which suggested the usefulness of using this analytical system for human identification as well as for forensic purposes.
Assuntos
Corantes Fluorescentes , Repetições Minissatélites , Paternidade , Polimorfismo Genético , Análise de Sequência de DNA/métodos , Automação , DNA/análise , Corantes Fluorescentes/química , Humanos , Indóis/química , Raios Infravermelhos , Estrutura Molecular , Técnicas de Amplificação de Ácido Nucleico , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Moldes GenéticosRESUMO
Anti-endomysial antibodies assayed by indirect immunofluorescence presently represent the most advanced in vitro method for diagnosing coeliac disease. While testing the serum of patients affected by coeliac disease for anti-endomysial antibodies, we noticed in some samples the presence of anti-smooth muscle antibodies. This led us to the hypothesis that anti-smooth muscle antibodies might mask the presence of anti-endomysial antibodies. The aim of our research was to develop a method that could confirm our hypothesis. Here we show that the AEA response can be effectively unmasked by increasingly diluting anti-smooth muscle antibodies-positive serum.
Assuntos
Autoanticorpos/sangue , Doença Celíaca/diagnóstico , Doença Celíaca/imunologia , Mucosa Intestinal/imunologia , Músculo Liso/imunologia , Técnica Indireta de Fluorescência para Anticorpo , Humanos , Técnicas de Diluição do Indicador , Lactente , Valor Preditivo dos Testes , Sensibilidade e Especificidade , Testes SorológicosRESUMO
We estimated the prevalence of celiac disease in children with juvenile chronic arthritis (JCA), using antiendomysium antibodies as the screening test to select patients for intestinal biopsy. We studied 119 children with JCA and found four patients with antiendomysium antibodies. In three of these patients (2.5%), intestinal biopsy revealed villous atrophy; in the fourth the intestinal mucosa was normal. We conclude that the prevalence of celiac disease is increased in patients with JCA.