Utilization of Multiple Real-Time PCR Assays for the Diagnosis of Bordetella spp. in Clinical Specimens

Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 943)

Abstract

Bordetella pertussis causes an upper respiratory infection in infants, adolescents, and adults. Diagnosis of pertussis, a vaccine-preventable disease, can be difficult, but recent implementation of real-time PCR assays in laboratories has hastened the ability of clinicians to make an accurate diagnosis. In this paper we describe the method of nasopharyngeal specimen collection, extraction of DNA, and real-time PCR assays that will allow the detection and identification of Bordetella spp. in clinical specimens.

References

  1. 1.
    Mattoo S, Cherry JD (2005) Molecular pathogenesis, epidemiology, and clinical manifestations of respiratory infections due to Bordetella pertussis and other Bordetella subspecies. Clin Microbiol Rev 18:326–382PubMedCrossRefGoogle Scholar
  2. 2.
    Goebel EM, Zhang X, Harvill ET (2009) Bordetella pertussis infection or vaccination substantially protects mice against B. bronchiseptica infection. PLoS One 4:e6778PubMedCrossRefGoogle Scholar
  3. 3.
    Tatti K, Wu K, Tondella L et al (2008) Development and evaluation of dual-target real-time polymerase chain reaction assays to detect Bordetella species. Diagn Microbiol Infect Dis 61:264–272PubMedCrossRefGoogle Scholar
  4. 4.
    CDC (2007) Outbreaks of respiratory illness mistakenly attributed to pertussis—New Hampshire, Massachusetts, and Tennessee, 2004–2006. MMWR 56:837–842Google Scholar
  5. 5.
    Tatti KM, Wu KH, Sanden GN et al (2006) Molecular diagnosis of Bordetella pertussis infection by evaluation of formalin-fixed tissue specimens. J Clin Microbiol 44:1074–1076PubMedCrossRefGoogle Scholar
  6. 6.
    Paddock CD, Sanden GN, Cherry JD et al (2008) Pathology and pathogenesis of fatal Bordetella pertussis infection in infants. Clin Infect Dis 47:28–38CrossRefGoogle Scholar
  7. 7.
    Qiagen (2007) QIAamp®DNA Mini and Blood Mini Handbook. Qiagen, Valencia, CAGoogle Scholar
  8. 8.
    Roche Applied Science (2008) MagNA Pure LC Total Nucleic Acid Isolation Kit Handbook. Roche Applied Science, MannheimGoogle Scholar
  9. 9.
    Espy MJ, Uhl JR, Sloan LM et al (2006) Real-time PCR in clinical microbiology: applications for routine laboratory testing. Clin Microbiol Rev 19:165–256PubMedCrossRefGoogle Scholar
  10. 10.
    Prophet EB, Mills B, Arrington JB, Sobin LH (1992) Laboratory methods in histotechnology. American Registry of Pathology, Washington, DCGoogle Scholar
  11. 11.
    Dundas N, Leos NK, Mitui M et al (2008) Comparison of automated nucleic acid extraction methods with manual extraction. J Mol Diagn 10:311–316PubMedCrossRefGoogle Scholar
  12. 12.
    Riffelman M, Schmetz J, Bock S et al (2007) Preparation of Bordetella pertussis DNA from respiratory samples for real-time PCR by commercial kits. Eur J Clin Microbiol Infect Dis 27:145–148CrossRefGoogle Scholar
  13. 13.
    Taranger J, Trollfors B, Lind L et al (1994) Environmental contamination leading to false-positive polymerase chain reaction for pertussis. Pediatr Infect Dis J 13:936–937PubMedGoogle Scholar
  14. 14.
    Tatti KM, Slade B, Patel M et al (2008) Real-time polymerase chain reaction detection of Bordetella pertussis DNA in acellular pertussis vaccines. Pediatr Infect Dis J 27:73–74PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2013

Authors and Affiliations

  1. 1.Division of Bacterial Diseases, Centers for Disease Control and Prevention, National Center for Immunization and Respiratory DiseasesMeningitis and Vaccine Preventable Diseases BranchAtlantaUSA

Personalised recommendations