title: Description of a multiplex Bordetella pertussis and Bordetella parapertussis LightCycler PCR assay with inhibition control.
abstract: While culture for Bordetella species is highly specific, sensitivity is extremely variable due to patient age, immunization status, antibiotic treatment, and specimen transport conditions. We evaluated a real-time multiplex PCR assay as an alternative to culture for the detection and differentiation of Bordetella pertussis and Bordetella parapertussis. The PCR conditions allowed the simultaneous detection of one B. pertussis organism and five B. parapertussis organisms per reaction. An inhibition control was incorporated into the assay. Of 163 total samples evaluated, 37 of 38 samples positive by either culture or direct fluorescent antibody testing (DFA) were also positive by PCR (97% sensitivity). Of 125 culture- or DFA-negative samples, 101 were also negative by PCR (81% specificity). The described multiplex assay is a rapid, sensitive, contamination-limiting, real-time PCR assay that controls for inhibition. The assay performs well using liquid or swab samples and from dried material on slides.
authors.affiliation: ARUP Institute for Clinical and Experimental Pathology, Salt Lake City, Utah, USA. cloudj1@aruplab.com
authors.forename: Joann L
authors.initials: JL
authors.lastname: Cloud
authors.forename: Weston C
authors.initials: WC
authors.lastname: Hymas
authors.forename: Arthur
authors.initials: A
authors.lastname: Turlak
authors.forename: Ann
authors.initials: A
authors.lastname: Croft
authors.forename: Udo
authors.initials: U
authors.lastname: Reischl
authors.forename: Judy A
authors.initials: JA
authors.lastname: Daly
authors.forename: Karen C
authors.initials: KC
authors.lastname: Carroll
mesh_terms: D000328:Adult; D001884:Bordetella / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D001885:Bordetella Infections / Q000175:diagnosis*; D001886:Bordetella pertussis / Q000235:genetics / Q000302:isolation & purification*; D002648:Child; D003470:Culture Media; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D014917:Whooping Cough / Q000175:diagnosis*
chemical_list: D003470:Culture Media