Real-time PCR assays of single-nucleotide polymorphisms defining the major Brucella clades.
J Clin Microbiol
; 46(1): 296-301, 2008 Jan.
Article
em En
| MEDLINE
| ID: mdl-18032628
ABSTRACT
Members of the genus Brucella are known worldwide as pathogens of wildlife and livestock and are the most common organisms of zoonotic infection in humans. In general, brucellae exhibit a range of host specificity in animals that has led to the identification of at least seven Brucella species. The genomes of the various Brucella species are highly conserved, which makes the differentiation of species highly challenging. However, we found single-nucleotide polymorphisms (SNPs) in housekeeping and other genes that differentiated the seven main Brucella species or clades and thus enabled us to develop real-time PCR assays based around these SNPs. Screening of a diverse panel of 338 diverse isolates with these assays correctly identified each isolate with its previously determined Brucella clade. Six of the seven clade-specific assays detected DNA concentrations of less than 10 fg, indicating a high level of sensitivity. This SNP-based approach places samples into a phylogenetic framework, allowing reliable comparisons to be made among the lineages of clonal bacteria and providing a solid basis for genotyping. These PCR assays provide a rapid and highly sensitive method of differentiating the major Brucella groups that will be valuable for clinical and forensic applications.
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Brucella
/
DNA Bacteriano
/
Reação em Cadeia da Polimerase
/
Técnicas de Tipagem Bacteriana
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Polimorfismo de Nucleotídeo Único
Tipo de estudo:
Diagnostic_studies
/
Evaluation_studies
Limite:
Animals
/
Humans
Idioma:
En
Revista:
J Clin Microbiol
Ano de publicação:
2008
Tipo de documento:
Article
País de afiliação:
Estados Unidos