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Evaluation of amplification targets for the specific detection of Bordetella pertussis using real-time polymerase chain reaction
- Source :
- The Canadian Journal of Infectious Diseases & Medical Microbiology, Canadian Journal of Infectious Diseases and Medical Microbiology, Vol 25, Iss 4, Pp 217-221 (2014)
- Publication Year :
- 2014
-
Abstract
- BACKGROUND:Bordetella pertussisinfections continue to be a major public health challenge in Canada. Polymerase chain reaction (PCR) assays to detectB pertussisare typically based on the multicopy insertion sequence IS481, which offers high sensitivity but lacks species specificity.METHODS: A novelB pertussisreal-time PCR assay based on the porin gene was tested in parallel with several previously published assays that target genes such as IS481,ptx-promoter, pertactin and a putative thialase. The assays were evaluated using a reference panel of common respiratory bacteria including differentBordetellaspecies and 107 clinical nasopharyngeal specimens. Discrepant results were confirmed by sequencing the PCR products.RESULTS: Analytical sensitivity was highest for the assay targeting the IS481element; however, the assay lacked specificity forB pertussisin the reference panel and in the clinical samples. False-positive results were also observed with assays targeting theptx-promoter and pertactin genes. A PCR assay based on the thialase gene was highly specific but failed to detect all reference strains ofB pertussis. However, a novel assay targeting the porin gene demonstrated high specificity forB pertussisboth in the reference panel and in clinical samples and, based on sequence-confirmed results, correctly predicted allB pertussis-positive cases in clinical samples. According to Probit regression analysis, the 95% detection limit of the new assay was 4 colony forming units/reaction.CONCLUSION: A novel porin assay forB pertussisdemonstrated superior performance and may be useful for improved molecular detection ofB pertussisin clinical specimens.
- Subjects :
- Microbiology (medical)
Bordetella pertussis
Specific detection
Pertactin gene
Ptx-promoter
Infectious and parasitic diseases
RC109-216
Biology
biology.organism_classification
Microbiology
Virology
IS481 element
QR1-502
law.invention
Porin gene
Infectious Diseases
Real-time polymerase chain reaction
law
B. pertussis
Original Article
Insertion sequence
Polymerase chain reaction
Real-time PCR
Subjects
Details
- ISSN :
- 17129532
- Volume :
- 25
- Issue :
- 4
- Database :
- OpenAIRE
- Journal :
- The Canadian journal of infectious diseasesmedical microbiology = Journal canadien des maladies infectieuses et de la microbiologie medicale
- Accession number :
- edsair.doi.dedup.....df4322c9eec52300cb215513823b56a0