title: Comparison of real-time PCR and pyrosequencing for typing Bordetella pertussis toxin subunit 1 variants.
abstract: We describe two newly developed methods for rapid typing of the pertussis toxin subunit 1 gene (ptxS1). A real-time PCR assay based on hybridization probes and a Pyrosequencing assay were developed and the specificity, sensitivity, cost, hands-on time and post-assay data processing were compared to Sanger sequencing. Both methods enabled discrimination of all four allelic variants, correctly identified all ptxS1 alleles of 143 strains tested and proved suitable for large-scale screening of B. pertussis strains.
authors.affiliation: Uppsala University Hospital, Department of Medical Sciences, Clinical Bacteriology, Uppsala, Sweden.
authors.forename: Martin
authors.initials: M
authors.lastname: Storm
authors.forename: Abdolreza
authors.initials: A
authors.lastname: Advani
authors.forename: Monica
authors.initials: M
authors.lastname: Pettersson
authors.forename: Hans O
authors.initials: HO
authors.lastname: Hallander
authors.forename: Kåre
authors.initials: K
authors.lastname: Bondeson
mesh_terms: D000483:Alleles; D001886:Bordetella pertussis / Q000145:classification / Q000235:genetics*; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D006801:Humans; D037342:Pertussis Toxin / Q000737:chemistry / Q000235:genetics*; D010567:Pertussis Vaccine / Q000235:genetics; D016133:Polymerase Chain Reaction; D020641:Polymorphism, Single Nucleotide; D017422:Sequence Analysis, DNA; D014917:Whooping Cough / Q000382:microbiology
chemical_list: D004269:DNA, Bacterial; D010567:Pertussis Vaccine; C083388:pertussis toxin, S1 subunit; D037342:Pertussis Toxin