The rationale and method for constructing internal control DNA used in pertussis polymerase chain reaction

Diagn Microbiol Infect Dis. 1998 Aug;31(4):517-23. doi: 10.1016/s0732-8893(98)00043-1.


The inclusion of an appropriate internal control DNA in polymerase chain reaction (PCR) is a rapid and simple method for the detection of PCR failure. Two PCR coamplification internal control DNAs (ICD I and ICD II) with the same primer-binding sequences as the target DNA for the detection of Bordetella pertussis and Bordetella parapertussis were produced using an overlap extension technique and a PCR MIMIC construction kit, respectively. The ICD II was further evaluated in a prospective clinical study in 360 patients with a clinical diagnosis of pertussis. From 360 nasopharyngeal swabs the internal control was positive in 318 (88%) samples, but was negative in 42 (12%). After phenol-chloroform extraction an additional 10 internal controls became positive. For the detection of PCR failure, the use of internal control DNA is highly recommended for PCR-based identification of B. pertussis and B. parapertussis organisms from nasopharyngeal swabs and aspirates.

MeSH terms

  • Bordetella / genetics
  • Bordetella / isolation & purification*
  • Bordetella pertussis / genetics
  • Bordetella pertussis / isolation & purification*
  • Child
  • DNA Primers
  • Evaluation Studies as Topic
  • False Negative Reactions
  • Humans
  • Nasopharynx / microbiology*
  • Polymerase Chain Reaction / methods*
  • Prospective Studies
  • Whooping Cough / diagnosis*


  • DNA Primers