Development of a multiplex real-time PCR assay for the detection of Bordetella pertussis and Bordetella parapertussis in a single tube reaction

Journal of Microbiological Methods
Sophie ArbefevillePatricia Ferrieri

Abstract

Pertussis is an infectious respiratory disease caused by the fastidious bacterium Bordetella pertussis, which may infect unvaccinated, previously vaccinated children, and adults in whom immunity has waned. Infants are at a particular risk for severe disease and complications. Bordetella parapertussis may cause a similar illness, however the symptoms are less severe and of shorter duration. Pertussis is a highly contagious disease and early diagnosis is essential. Studies have shown that PCR is 2-4 times more likely than culture to detect Bordetella pertussis. We developed a multiplex, real-time PCR assay using analyte-specific reagent (ASR) primers and probes dispensed in a convenient lyophilized bead format that targeted the multi-copy insertion sequences IS481 and IS1001 of B. pertussis and B. parapertussis, respectively. These specific ASRs were used in conjunction with Cepheid Smartmix. Included in the ASRs is a competitive internal control to evaluate the performance of the PCR reaction. After DNA extraction, amplification and detection were done on the Smart Cycler System, which performs integrated amplification and detection automatically in a single step. Specificity of the assay was confirmed using multiple distinct ba...Continue Reading

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Citations

Nov 3, 2015·International Journal of Infectious Diseases : IJID : Official Publication of the International Society for Infectious Diseases·María Esther CastilloJuana Del Valle Mendoza
Oct 16, 2020·Journal of Clinical Microbiology·Siu-Kei ChowTed E Schutzbank
Mar 9, 2021·Food and Waterborne Parasitology·Alexandre D T CostaMarco A Krieger

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