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Trangoni,Marcos D.; Gioffré,Andrea K.; Cerón Cucchi,María E.; Caimi,Karina C.; Ruybal,Paula; Zumárraga,Martín J.; Cravero,Silvio L.. |
In this study, we developed new sets of primers to detect Brucella spp. and M. avium subsp. paratuberculosis (MAP) through isothermal amplification. We selected a previously well-characterized target gene, bscp31, specific for Brucella spp. and IS900 for MAP. The limits of detection using the loop-mediated isothermal amplification (LAMP) protocols described herein were similar to those of conventional PCR targeting the same sequences. Hydroxynaphtol blue and SYBR GreenTM allowed direct naked-eye detection with identical sensitivity as agarose gel electrophoresis. We included the LAMP-based protocol in a rapid identification scheme of the respective pathogens, and all tested isolates were correctly identified within 2 to 3 h. In addition, both protocols... |
Tipo: Info:eu-repo/semantics/article |
Palavras-chave: Loop-mediated isothermal amplification; Molecular typing; Brucellosis; Paratuberculosis. |
Ano: 2015 |
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822015000200619 |
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Gioffré,Andrea; Correa Muñoz,Magnolia; Alvarado Pinedo,María F.; Vaca,Roberto; Morsella,Claudia; Fiorentino,María Andrea; Paolicchi,Fernando; Ruybal,Paula; Zumárraga,Martín; Travería,Gabriel E.; Romano,María Isabel. |
Multiple-locus variable number-tandem repeat analysis (MLVA) of Mycobacterium avium subspecies paratuberculosis (MAP) isolates may contribute to the knowledge of strain diversity in Argentina. Although the diversity of MAP has been previously investigated in Argentina using IS900-RFLP, a small number of isolates were employed, and a low discriminative power was reached. The aim of the present study was to test the genetic diversity among MAP isolates using an MLVA approach based on 8 repetitive loci. We studied 97 isolates from cattle, goat and sheep and could describe 7 different patterns: INMV1, INMV2, INMV11, INMV13, INMV16, INMV33 and one incomplete pattern. INMV1 and INMV2 were the most frequent patterns, grouping 76.3% of the isolates. We were also... |
Tipo: Info:eu-repo/semantics/article |
Palavras-chave: Paratuberculosis; Molecular typing; MIRU-VNTR; MLVA. |
Ano: 2015 |
URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822015000200557 |
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