Development of real-time PCR assay for differential detection of Bordetella bronchiseptica and Bordetella parapertussis
Language English Country United States Media print-electronic
Document type Evaluation Study, Journal Article, Research Support, Non-U.S. Gov't
PubMed
24525142
DOI
10.1016/j.diagmicrobio.2013.12.020
PII: S0732-8893(14)00013-3
Knihovny.cz E-resources
- Keywords
- Bordetella bronchiseptica, Bordetella parapertussis, Diagnosis, Real-time PCR,
- MeSH
- Bordetella bronchiseptica genetics isolation & purification MeSH
- Bordetella parapertussis genetics isolation & purification MeSH
- Molecular Diagnostic Techniques methods MeSH
- Diagnosis, Differential MeSH
- DNA, Bacterial chemistry genetics MeSH
- Bordetella Infections diagnosis microbiology MeSH
- Real-Time Polymerase Chain Reaction methods MeSH
- Humans MeSH
- Molecular Sequence Data MeSH
- Sequence Analysis, DNA MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
- Evaluation Study MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- DNA, Bacterial MeSH
Bordetella parapertussis is a causative agent of whooping cough in humans, and B. bronchiseptica is causing wide variety of respiratory infections in mammals, including humans. Specific diagnostic tests are not currently available. Our first objective was to develop a real-time PCR test for the specific detection of B. bronchiseptica based on the previously described end-point PCR, targeting an intergenomic sequence of the fla gene locus, but it has not been reached. However, there is cross-reactivity between B. parapertussis and B. bronchiseptica. Therefore, the targeted region of several clinical isolates of both species was sequenced, and alignment of the sequences allowed the development of a 2-step real-time PCR assay. The first PCR assay detected the DNA of all clinical isolates of both B. bronchiseptica and B. parapertussis tested. The second PCR assay detected only the DNA of B. parapertussis clinical isolates, thereby allowing discrimination between B. parapertussis and B. bronchiseptica.
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