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LAMP technology: Rapid identification of Brucella and Mycobacterium avium subsp. paratuberculosis BJM
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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Rapid, specific, and sensitive detection of the ureR_1 gene in Klebsiella pneumoniae by loop-mediated isothermal amplification method BJMBR
Li,Chao; Fu,Gongyu; Shi,Yaoqiang; Zhang,A-Mei; Xia,Xueshan; Fang,Yue; Mao,Xiaoqin; Jiang,Jie; Song,Yuzhu; Yang,Guangying.
Klebsiella pneumoniae is one of the main pathogenic bacteria that causes nosocomial infections, such as pneumonia, urinary tract infection, and sepsis. Therefore, the rapid and accurate detection of K. pneumoniae is important for the timely treatment of infectious patients. This study aimed to establish a loop-mediated isothermal amplification (LAMP) method for the rapid and sensitive detection of K. pneumoniae-specific gene ureR_1 (Gene ID: 11847803). The ureR_1 gene was obtained through local and online BLAST, and the specific primers were designed for its detection. Positive reactions were observed on all 140 K. pneumoniae clinical isolates while all the 82 non-K. pneumoniae clinical isolates were negative. Plasmids with the specific gene and the mouse...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Loop-mediated isothermal amplification; Polymerase chain reaction; Klebsiella pneumoniae; Novel specific gene; Specific; Sensitive method.
Ano: 2019 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2019000300609
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The development of a loop-mediated isothermal amplification assay for rapid and sensitive detection of abalone herpesvirus DNA ArchiMer
Chen, Meilin; Kuo, S. T.; Renault, Tristan; Chang, P. H..
A loop-mediated isothermal amplification (LAMP) assay was developed for the detection of abalone herpesvirus DNA. Two pairs of primers were designed, based on the sequence of the DNA polymerase gene of abalone herpesvirus. The reaction temperature and time were optimized to 63 °C and 60 min, respectively. LAMP amplicons were analyzed by 2% agarose gel electrophoresis or by visual inspection of a colour change emitted by fluorescent dye. The method developed was specific for the detection of abalone herpesvirus, without cross-reactions with other tested herpesviruses including ostreid herpesvirus 1 (OsHV-1), European eel herpesvirus, koi herpesvirus (KHV) and an avian herpesvirus. The LAMP assay was 100 folds more sensitive than a conventional PCR and 10...
Tipo: Text Palavras-chave: Herpesvirus; Abalone; Haliotis diversicolor supertexta; Polymerase chain reaction; Loop-mediated isothermal amplification; SYBR green PCR.
Ano: 2014 URL: https://archimer.ifremer.fr/doc/00166/27690/25882.pdf
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Yersinia pestis detection by loop-mediated isothermal amplification combined with magnetic bead capture of DNA BJM
Feng,Na; Zhou,Yazhou; Fan,Yanxiao; Bi,Yujing; Yang,Ruifu; Zhou,Yusen; Wang,Xiaoyi.
ABSTRACT We developed a loop-mediated isothermal amplification (LAMP) assay for the detection of Y. pestis by targeting the 3a sequence on chromosome. All 11 species of the genus Yersinia were used to evaluate the specificity of LAMP and PCR, demonstrating that the primers had a high level of specificity. The sensitivity of LAMP or PCR was 2.3 or 23 CFU for pure culture, whereas 2.3 × 104 or 2.3 × 106 CFU for simulated spleen and lung samples. For simulated liver samples, the sensitivity of LAMP was 2.3 × 106 CFU, but PCR was negative at the level of 2.3 × 107 CFU. After simulated spleen and lung samples were treated with magnetic beads, the sensitivity of LAMP or PCR was 2.3 × 103 or 2.3 × 106 CFU, whereas 2.3 × 105 or 2.3 × 107 CFU for magnetic...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Plague; Yersinia pestis; Loop-mediated isothermal amplification; Magnetic beads.
Ano: 2018 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822018000100128
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