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. 2004;18(5):265-70.
doi: 10.1002/jcla.20035.

Rapid detection of Bordetella pertussis by real-time PCR using SYBR green I and a LightCycler instrument

Affiliations

Rapid detection of Bordetella pertussis by real-time PCR using SYBR green I and a LightCycler instrument

S K Poddar. J Clin Lab Anal. 2004.

Abstract

A polymerase chain reaction (PCR) assay in real-time for detection of B. pertussis using SYBR green I as the reporter fluorophore and LightCycler instrument (a thermocycler coupled to a fluorescence detection device) was established and evaluated. The amplified amplicon using series diluted control prototype strain (ATCC strain #9797) of B. pertussis was analyzed for the fluorescent melting profile, and melting temperature (Tm) was determined. When examined, amplicons using a representative set of clinical isolates of B. pertussis were found to have the same Tm value (86 +/- 0.5 degrees C, the specificity parameter of detection) as the control prototype strain as expected. Amplified product was also analyzed and detected by agarose gel electrophoresis. The detection limit by fluorescent profile and Tm analysis was 10-fold better than that detected by agarose gel analysis.

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References

    1. Cherry JD. Historical review of pertussis and the classical vaccine. J Infect Dis 1996;174(suppl 3):S259–S263. - PubMed
    1. Stojanov S, Liese J, Belohradsky BH. Hospitalization and complications in children under 2 years of age with Bordetella pertussis infection. Infection 2000;28:106–110. - PubMed
    1. Khetsuriani NK, Bisgard DR, Prevots M, et al. Pertussis outbreak in an elementary school with high vaccination coverage. Pediatr Infect Dis J 2001;20:1108–1112. - PubMed
    1. He Q, Viljanen MK, Arvilommi H, Aittanen B, Mertsola J. Whooping cough caused by Bordetella pertussis and Bordetella parapertussis in an immunized population. JAMA 1998;280:635–637. - PubMed
    1. SenZilet LC, Halperin SA, Spika M, Alagaratnam A, Morris A, Smith B. Pertussis is a frequent cause of prolonged cough illness in adults and adolescents. Clin Infect Dis 2001;32:1691–1697. - PubMed

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