Comparison of real-time PCR and conventional hemi-nested PCR for the detection of Bordetella pertussis in nasopharyngeal samples

Clin Microbiol Infect. 2003 Jul;9(7):746-9. doi: 10.1046/j.1469-0691.2003.00601.x.


We directly compared a conventional hemi-nested PCR assay with a real-time (LightCycler) PCR assay for the detection of Bordetella pertussis in nasopharyngeal samples. Of the 152 samples tested, 49 (32%) were positive by first-round conventional PCR, 56 (37%) were positive by the hemi-nested PCR assay, and 39 (26%) were positive by the real-time assay. All samples testing positive with the real-time assay were also positive by the hemi-nested assay (both first- and second-round PCR), and all culture-positive samples tested positive by both PCR assays. These results suggest that the hemi-nested assay is more sensitive than the real-time assay for detecting B. pertussis.

Publication types

  • Comparative Study

MeSH terms

  • Adolescent
  • Adult
  • Aged
  • Bordetella pertussis / genetics
  • Bordetella pertussis / isolation & purification*
  • Child
  • Child, Preschool
  • Humans
  • Infant
  • Infant, Newborn
  • Middle Aged
  • Nasopharyngeal Diseases / diagnosis
  • Nasopharyngeal Diseases / microbiology
  • Polymerase Chain Reaction / methods*
  • Whooping Cough / diagnosis*