Detection of Bordetella pertussis in clinical specimens by PCR and a microtiter plate-based DNA hybridization assay.

Journal of Clinical Microbiology
S NelsonL Dyster

Abstract

In order to improve detection of Bordetella pertussis in nasopharyngeal aspirates (NPAs) in our laboratory, a PCR-based assay was optimized, and a study was designed (i) to compare results obtained by PCR to those obtained by culture and (ii) to evaluate a novel microtiter plate-based DNA hybridization assay (PCR-plate) by comparing it to agarose gel electrophoresis (PCR-gel) for detection of the PCR product. DNA for the PCR was extracted with a guanidine thiocyanate buffer and used in a PCR mixture containing primers directed against a reiterated gene sequence in B. pertussis (Q. He, J. Mertsola, H. Soini, M. Skurnik, O. Ruuskanen, and M. K. Viljanen, J. Clin, Microbiol. 31:642-645, 1993). Of 96 NPAs submitted from a targeted study group, 23 were positive by culture, 27 were positive by PCR-gel, and 31 were positive by PCR-plate. All culture-positive specimens were also positive by PCR. Of nine patients with culture-negative-PCR-positive results, six had discharge diagnoses of pertussis. Thus, PCR with plate-based product detection is a sensitive method for the laboratory detection of B. pertussis in NPAs. Additional advantages of the plate assay include rapidity, objectivity in reading results, specificity, and the capability...Continue Reading

References

Mar 1, 1994·Journal of Clinical Microbiology·Z LiM J Brennan

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Citations

Feb 16, 2005·European Journal of Clinical Microbiology & Infectious Diseases : Official Publication of the European Society of Clinical Microbiology·W B ParkK W Choe
Jun 30, 2009·The Pediatric Infectious Disease Journal·Valerie WatersScott A Halperin
Oct 14, 2005·The New England Journal of Medicine·Joel I WardUNKNOWN APERT Study Group
May 8, 2001·Clinical Infectious Diseases : an Official Publication of the Infectious Diseases Society of America·J S TanJ E Stout
Sep 12, 2015·Clinical Microbiology Reviews·Anneke van der ZeeFrits R Mooi
May 6, 2005·Journal of Clinical Microbiology·Stéphanie HerweghRené J Courcol
May 9, 1998·Journal of Clinical Microbiology·S NelsonP Quinn
Aug 17, 1999·Journal of Clinical Microbiology·M J LoeffelholzM J Gilchrist

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