Bordetella pertussis PCR: Simultaneous targeting of signature sequences

Xuan Qin, David K. Turgeon, Brian P. Ingersoll, Peter W. Monsaas, Christina J. Lemoine, Treva Tsosie, Lynn O. Stapp, Patrick M. Abe

Research output: Contribution to journalArticlepeer-review

19 Scopus citations

Abstract

The absence of analytical controls for polymerase chain reaction (PCR)-based diagnostic tests for Bordetella pertussis limits their clinical utility. In this study, multiplex PCR simultaneously targeted two specific Bordetella pertussis sequences, the chromosomal repeated insertion sequence IS481 (IS) and the pertussis toxin promoter region (PT). A multi-target hybridization-EIA (Hyb-EIA) method in a 96-well microtiter-plate format was used to detect amplicons. Forty-seven (15%) of the 318 nasopharygeal specimens tested positive for at least one DNA target of B. pertussis by PCR, including the 10 known positive samples by culture and/or direct fluorescent antibody (DFA). Forty-six of the 47 PCR positive samples were considered positive for B. pertussis using the consensus interpretation criteria. Simultaneous detection of multiple chromosomal regions may identify false-positive and -negative results due to analytical variations or potential sequence polymorphism, and uncover a wider range of pathogenic strains.

Original languageEnglish (US)
Pages (from-to)269-275
Number of pages7
JournalDiagnostic Microbiology and Infectious Disease
Volume43
Issue number4
DOIs
StatePublished - 2002
Externally publishedYes

Keywords

  • Analytical control
  • Multiplex PCR
  • Sequence polymorphism

ASJC Scopus subject areas

  • Microbiology (medical)
  • Infectious Diseases

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