RESUMO
The results of MLPA test using serum and filter paper eluate have been compared in this paper. Testing 64 patient samples at 1:32 dilution, 31 were negative by both serum and eluate, 20 were positive by both, six were positive only by serum and one was positive only eluate. In six other cases eluate gave equivocal results while serum result was clearly positive. Some eluates negative at 1:32 dilution gave weak positive agglutination at 1:16 dilution.
Assuntos
Hanseníase/sangue , Mycobacterium leprae , Testes de Aglutinação/métodos , Coleta de Amostras Sanguíneas/métodos , Humanos , Fitas ReagentesRESUMO
The gene for Aspergillus niger glucose oxidase (EC 1.1.3.4) has been cloned from both cDNA and genomic libraries using oligonucleotide probes derived from the amino acid sequences of peptide fragments of the enzyme. The mature enzyme consists of 583 amino acids and is preceded by a 22-amino acid presequence. No intervening sequences are found within the coding region. The enzyme contains 3 cysteine residues and 8 potential sites for N-linked glycosylation. The protein shows 26% identity with alcohol oxidase of Hansenuela polymorpha, and the N terminus has a sequence homologous with the AMP-binding region of other flavoenzymes such as p-hydroxybenzoate hydroxylase and glutathione reductase. Recombinant yeast expression plasmids have been constructed containing a hybrid yeast alcohol dehydrogenase II-glyceraldehyde-3-phosphate dehydrogenase promoter, either the yeast alpha-factor pheromone leader or the glucose oxidase presequence, and the mature glucose oxidase coding sequence. When transformed into yeast, these plasmids direct the synthesis and secretion of between 75 and 400 micrograms/ml of active glucose oxidase. Analysis of the yeast-derived enzymes shows that they are of comparable specific activity and have more extensive N-linked glycosylation than the A. niger protein.
Assuntos
Aspergillus niger/genética , Clonagem Molecular , Genes Fúngicos , Glucose Oxidase/genética , Saccharomyces cerevisiae/genética , Sequência de Aminoácidos , Aspergillus niger/enzimologia , Sequência de Bases , Glucose Oxidase/biossíntese , Glucose Oxidase/metabolismo , Cinética , Dados de Sequência Molecular , Fragmentos de Peptídeos/isolamento & purificação , Plasmídeos , Proteínas Recombinantes/biossíntese , Proteínas Recombinantes/metabolismo , Mapeamento por Restrição , Saccharomyces cerevisiae/enzimologia , Homologia de Sequência do Ácido NucleicoRESUMO
Response of Mycobacterium habana sensitized mouse cells against habanin (sonicated extract) lepromin (Dharmendra) and tuberculin has been studied in the leucocyte migration inhibition test. The homologous antigenic response has evoked maximum inhibition as compared to heterologous antigens but close antigenic association has also been observed with lepromin and tuberculin. With human cells from leprosy patients these antigens have evoked analogous response indicating cross immunogenicity of M. habana with leprosy sensitized cells.