Back to Search
Start Over
Yersinia pestis detection by loop-mediated isothermal amplification combined with magnetic bead capture of DNA
- Source :
- Brazilian Journal of Microbiology, Brazilian Journal of Microbiology v.49 n.1 2018, Sociedade Brasileira de Microbiologia (SBM), instacron:SBM, Brazilian Journal of Microbiology, Vol 49, Iss 1, Pp 128-137, Brazilian Journal of Microbiology, Volume: 49, Issue: 1, Pages: 128-137, Published: MAR 2018
- Publication Year :
- 2017
- Publisher :
- Elsevier, 2017.
-
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 bead-treated liver samples. These results indicated that some components in the tissues could inhibit LAMP and PCR, and liver tissue samples had a stronger inhibition to LAMP and PCR than spleen and lung tissue samples. LAMP has a higher sensitivity than PCR, and magnetic bead capture of DNAs could remarkably increase the sensitivity of LAMP. LAMP is a simple, rapid and sensitive assay suitable for application in the field or poverty areas.
- Subjects :
- 0301 basic medicine
DNA, Bacterial
genetic structures
Yersinia pestis
030106 microbiology
Loop-mediated isothermal amplification
lcsh:QR1-502
Spleen
Biology
Microbiology
Polymerase Chain Reaction
Sensitivity and Specificity
lcsh:Microbiology
03 medical and health sciences
chemistry.chemical_compound
Magnetics
Liver tissue
medicine
Humans
DNA Primers
Plague
Magnetic beads
Immunomagnetic Separation
biology.organism_classification
Genus Yersinia
Molecular biology
eye diseases
030104 developmental biology
medicine.anatomical_structure
chemistry
Magnetic bead
sense organs
Lung tissue
Nucleic Acid Amplification Techniques
DNA
Research Paper
Subjects
Details
- Language :
- English
- ISSN :
- 16784405 and 15178382
- Volume :
- 49
- Issue :
- 1
- Database :
- OpenAIRE
- Journal :
- Brazilian Journal of Microbiology
- Accession number :
- edsair.doi.dedup.....5bd0b3bd7e498379fb1618f51594d765