Laboratory diagnosis of pertussis infections: the role of PCR and serology

1,2 Respiratory and Systemic Infection Laboratory 1 and Special Projects Laboratory 2 , Specialist and Reference Microbiology Division, Health Protection Agency, 61 Colindale Avenue, London NW9 5HT, UK 3 Health Protection Agency, Immunization Division, Communicable Disease Surveillance Centre, 61 Co...

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Veröffentlicht in:Journal of medical microbiology 2004-06, Vol.53 (6), p.519-525
Hauptverfasser: Fry, Norman K, Tzivra, Oceanis, Li, Y. Ting, McNiff, Anthony, Doshi, Nivedita, Maple, P. A.Christopher, Crowcroft, Natasha S, Miller, Elizabeth, George, Robert C, Harrison, Timothy G
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Sprache:eng
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Zusammenfassung:1,2 Respiratory and Systemic Infection Laboratory 1 and Special Projects Laboratory 2 , Specialist and Reference Microbiology Division, Health Protection Agency, 61 Colindale Avenue, London NW9 5HT, UK 3 Health Protection Agency, Immunization Division, Communicable Disease Surveillance Centre, 61 Colindale Avenue, London NW9 5HT, UK Correspondence Norman K. Fry Norman.Fry{at}HPA.org.uk Received February 4, 2004 Accepted February 12, 2004 This study reports on practical laboratory aspects of pertussis diagnosis. PCR assays were applied to respiratory specimens obtained during a large study of infants (less than 5 months old) admitted to paediatric intensive care units ( n = 122), children (less than 15 years old) admitted to paediatric wards ( n = 16) and their household contacts ( n = 320). Estimation of antibodies to pertussis toxin and culture for Bordetella pertussis were attempted on specimens from the same patients, where available, and the overall utility of the diagnostic PCR assays was assessed by comparison to these results. A PCR assay for the human mitochondrial cytochrome oxidase (HMCO) gene was used for quality control of the extracted samples and an internal process control (IPC) was included in each sample to test for PCR inhibition. Four of 458 samples were considered unsuitable (three HMCO negative, one IPC negative) and excluded from further analyses. Positive PCR results were considered valid if they were either (i) positive for both of two B. pertussis gene targets (pertussis toxin S1 promoter and the insertion element IS 481 ), i.e. consensus PCR positive, or (ii) repeatably positive in only one assay. Using these criteria, 52 of 454 (11.5 %) samples were considered as PCR positive for B. pertussis . Six of 356 samples were culture-positive for B. pertussis , 1/88 infants, 3/14 children and 2/254 contacts, giving an overall isolation rate of 1.7 %. Using these data, PCR gave an almost fivefold increase in diagnostic yield compared with culture (McNemar's test; P < 0.0001). Sera from 9/111 infants, 5/10 children and 14/210 contacts were positive. Serology and PCR results showed a high level of agreement (113/121) for infants and children. PCR demonstrated a significant improvement in diagnostic yield over culture. Serological testing also resulted in a significant increase in diagnostic yield compared to culture alone. PCR is a useful technique, but validity of results must be assured by careful control. Rapid diagnosis of B. pertussis
ISSN:0022-2615
1473-5644
DOI:10.1099/jmm.0.45624-0