Comparison of real-time PCR and pyrosequencing for typing Bordetella pertussis toxin subunit 1 variants

• 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.

Nyckelord

Alleles

Bordetella pertussis/classification/*genetics

DNA; Bacterial/chemistry/genetics

Humans

Pertussis Toxin/chemistry/*genetics

Pertussis Vaccine/genetics

Polymerase Chain Reaction

Polymorphism; Single Nucleotide

Sequence Analysis; DNA

Whooping Cough/microbiology

MEDICINE

MEDICIN

Publikations- och innehållstyp

ref (ämneskategori)

art (ämneskategori)