INVESTIGADORES
HOZBOR Daniela Flavia
artículos
Título:
Detection of Bordetella bronchiseptica by polymerase chain reaction
Autor/es:
HOZBOR D; FOUQUE F; GUISO N
Revista:
RESEARCH IN MICROBIOLOGY
Editorial:
ELSEVIER SCIENCE BV
Referencias:
Lugar: Amsterdam; Año: 1999 p. 333 - 341
ISSN:
0923-2508
Resumen:
Polymerase chain reaction (PCR) assays were developed that enabled not
only discriminative detection of three Bordetella species, B. pertussis,
B. parapertussis, and B. bronchiseptica (Bspp PCR), but also specific
detection of B. bronchiseptica (Bb PCR). An upstream sequence of the
flagellin gene was used as a target DNA region. This sequence contained
differences in B. pertussis, B. parapertussis, and B. bronchiseptica
DNA. These species could then be differentiated using two different sets
of primers, Bspp and Bb. When oligonucleotide Bspp primers were used,
PCR products were obtained from the three species of Bordetella. A
fragment of the expected size (164 bp) was amplified using B.
bronchiseptica and B. parapertussis DNA, but a fragment with a distinct
molecular weight was amplified with B. pertussis DNA (195 bp). This Bspp
PCR was specific and sensitive, but it could not differentiate between
B. parapertussis and B. bronchiseptica. When Bb primers were used, a
237-bp PCR product was detected only from B. bronchiseptica DNA. No PCR
products were identified after Bb PCR amplification of DNAs either from
B. parapertussis isolates or B. pertussis isolates, nor from other
respiratory pathogen DNAs tested. This second PCR assay had a
sensitivity limit of less than 10 organisms of B. bronchiseptica after
detection with a specific probe. The specificity and the sensitivity of
the fla PCR assay were evaluated with purified DNA, as was its capacity
for detecting the bacteria in human clinical samples and in lungs of
infected mice.