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Journal of Clinical Microbiology, August 2002, p. 2908-2912, Vol. 40, No. 8
0095-1137/02/$04.00+0     DOI: 10.1128/JCM.40.8.2908-2912.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Diagnosis of Community-Acquired Pertussis Infection: Comparison of Both Culture and Fluorescent-Antibody Assays with PCR Detection Using Electrophoresis or Dot Blot Hybridization

Jairam R. Lingappa,^1* William Lawrence,² Sheyla West-Keefe,¹ Romesh Gautom,² and Brad T. Cookson^1,3

Department of Laboratory Medicine,¹ Department of Microbiology, University of Washington,³ Washington State Public Health Laboratories, Seattle, Washington²

Received 29 January 2002/ Returned for modification 5 April 2002/ Accepted 13 May 2002

Diagnosis of Bordetella pertussis infection has been difficult due to the low sensitivity of culture. PCR tests have been shown to be more sensitive than culture, but the reported sensitivity of PCR is variable. We evaluated PCR product detection by using either agarose gel electrophoresis (PCR-gel) or dot blot hybridization with ³²P-labeled oligonucleotide probes, and we compared these methods to both culture and direct fluorescent-antibody (DFA) assays with microscopy for the detection of pertussis. This was done with 225 nasopharyngeal swab specimens collected in community clinic settings. The multiplexed PCR amplified the multiply repeated IS481 B. pertussis sequence and a sequence from the human globin gene as a positive control for specimen adequacy. Of 225 specimens, 179 were judged to be adequate for PCR analysis. Among the adequate specimens, 9, 4, and 10 were culture, DFA, and PCR-gel positive, respectively. The sensitivity of PCR-gel versus culture was 89% while the sensitivity of culture versus PCR-gel was 80%. DFA had the lowest sensitivity. Thirty specimens were positive by PCR with dot blot hybridization; no negative control specimens showed a signal above the background. Among the 79 (44%) adequate specimens with clinical data available, the rates of reported cough or persistent cough were similar for persons who were pertussis positive by each assay. The IS481 PCR, with either electrophoresis or dot blot hybridization, is a sensitive assay; however, at this time it cannot completely replace culture without an overall loss in sensitivity for the detection of pertussis. Further study is required to understand the clinical significance of B. pertussis PCR products detected by dot blot hybridization alone.

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* Corresponding author. Present address: Division of Viral and Rickettsial Diseases, National Center for Infectious Diseases, Centers for Disease Control and Prevention, Mail Stop A-34, 1600 Clifton Rd., Atlanta, GA 30333. Phone: (404) 639-3596. Fax: (404) 639-1307. E-mail: JWL8{at}cdc.gov.

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Journal of Clinical Microbiology, August 2002, p. 2908-2912, Vol. 40, No. 8
0095-1137/02/$04.00+0     DOI: 10.1128/JCM.40.8.2908-2912.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

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