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Identification of Bordetella pertussis in nasopharyngeal swabs by PCR amplification of a region of the adenylate cyclase gene.
- Source :
-
Journal of medical microbiology [J Med Microbiol] 1993 Feb; Vol. 38 (2), pp. 140-4. - Publication Year :
- 1993
-
Abstract
- The polymerase chain reaction (PCR) was used to amplify a 522-bp region of the adenylate cyclase toxin (cyaA) gene of Bordetella pertussis. As few as 100 cfu from a suspension of B. pertussis could be detected by this procedure when the amplified PCR product was detected by ethidium bromide staining of agarose gels. However, simulated clinical specimens, prepared from swabs impregnated with known numbers of B. pertussis cells, only yielded a positive reaction with > or = 10(4) cfu. Hybridisation of a Southern blot of the PCR products from the swab samples with a cya-specific probe gave a positive reaction with as few as 8 cfu, but the hybridisation signal was uniformly weak with fewer than 10(4) cfu. Nevertheless, three of 13 nasopharyngeal swabs, taken from suspected clinically defined cases of whooping cough and stored frozen for up to 18 months, gave a positive PCR reaction.
- Subjects :
- Bordetella pertussis enzymology
Bordetella pertussis genetics
DNA, Bacterial isolation & purification
Evaluation Studies as Topic
Humans
Nucleic Acid Hybridization
Virulence Factors, Bordetella isolation & purification
Whooping Cough microbiology
Adenylate Cyclase Toxin
Bordetella pertussis isolation & purification
Nasal Cavity microbiology
Polymerase Chain Reaction
Virulence Factors, Bordetella genetics
Subjects
Details
- Language :
- English
- ISSN :
- 0022-2615
- Volume :
- 38
- Issue :
- 2
- Database :
- MEDLINE
- Journal :
- Journal of medical microbiology
- Publication Type :
- Academic Journal
- Accession number :
- 8429539
- Full Text :
- https://doi.org/10.1099/00222615-38-2-140