LAMP technology: Rapid identification of Brucella and Mycobacterium avium subsp. paratuberculosis
Braz. j. microbiol
; 46(2): 619-626, Apr-Jun/2015. tab, graf
Article
in English
| LILACS
| ID: lil-749730
Responsible library:
BR1.1
ABSTRACT
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 were suitable for specifically identifying the respective pathogens; in the case of Brucella, it also allowed the identification of all the biovars tested. We conclude that LAMP is a suitable rapid molecular typing tool that could help to shorten the time required to identify insidious bacteria in low-complexity laboratories, mainly in developing countries.
Full text:
Available
Collection:
International databases
Health context:
Neglected Diseases
Health problem:
Neglected Diseases
/
Zoonoses
Database:
LILACS
Main subject:
Brucella
/
Mycobacterium avium subsp. paratuberculosis
/
Nucleic Acid Amplification Techniques
/
Molecular Diagnostic Techniques
Type of study:
Diagnostic study
/
Evaluation study
/
Prognostic study
Limits:
Animals
/
Humans
Language:
English
Journal:
Braz. j. microbiol
Journal subject:
Microbiology
Year:
2015
Document type:
Article
/
Project document
Affiliation country:
Argentina
Institution/Affiliation country:
National Agricultural Technology Institute/AR