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Optimization of randomly amplified polymorphic DNA-polymerase chain reaction for molecular typing of Salmonella enterica serovar Typhi
- Source :
- Revista da Sociedade Brasileira de Medicina Tropical. 37:143-147
- Publication Year :
- 2004
- Publisher :
- FapUNIFESP (SciELO), 2004.
-
Abstract
- Optimization of the RAPD reaction for characterizing Salmonella enterica serovar Typhi strains was studied in order to ensure the reproducibility and the discriminatory power of this technique. Eight Salmonella serovar Typhi strains isolated from various regions in Brazil were examined for the fragment patterns produced using different concentrations of DNA template, primer, MgCl2 and Taq DNA polymerase. Using two different low stringency thermal cycle profiles, the RAPD fingerprints obtained were compared. A set of sixteen primers was evaluated for their ability to produce a high number of distinct fragments. We found that variations associated to all of the tested parameters modified the fingerprinting patterns. For the strains of Salmonella enterica serovar Typhi used in this experiment, we have defined a set of conditions for RAPD-PCR reaction, which result in a simple, fast and reproducible typing method.
- Subjects :
- DNA, Bacterial
Microbiology (medical)
Serotype
Salmonella
DNA polymerase
medicine.disease_cause
Microbiology
medicine
Humans
Typing
biology
Reproducibility of Results
Salmonella typhi
bacterial infections and mycoses
DNA Fingerprinting
Molecular biology
Bacterial Typing Techniques
Random Amplified Polymorphic DNA Technique
RAPD
Infectious Diseases
Salmonella enterica serovar Typhi
biology.protein
Parasitology
Primer (molecular biology)
Chain reaction
Brazil
Subjects
Details
- ISSN :
- 00378682
- Volume :
- 37
- Database :
- OpenAIRE
- Journal :
- Revista da Sociedade Brasileira de Medicina Tropical
- Accession number :
- edsair.doi.dedup.....b85060e07cc9f0c4b7b5e350f58fc93e