1887

Abstract

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 ( = 122), children (less than 15 years old) admitted to paediatric wards ( = 16) and their household contacts ( = 320). Estimation of antibodies to pertussis toxin and culture for 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 gene targets (pertussis toxin S1 promoter and the insertion element IS), 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 . Six of 356 samples were culture-positive for , 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; < 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 infection particularly in infants by PCR, together with serological assays, can enhance surveillance systems for pertussis in all age groups.

Loading

Article metrics loading...

/content/journal/jmm/10.1099/jmm.0.45624-0
2004-06-01
2024-04-19
Loading full text...

Full text loading...

/deliver/fulltext/jmm/53/6/JM530609.html?itemId=/content/journal/jmm/10.1099/jmm.0.45624-0&mimeType=html&fmt=ahah

References

  1. Anonymous. 2003 Investigation of specimens for Bordetella species . Health Protection Agency Standard Operating Procedure (BSOP 6i5.1) London, UK:
    [Google Scholar]
  2. Cadieux N., Lebel P., Brousseau R. 1993; Use of a triplex polymerase chain reaction for the detection and differentiation of Mycoplasma pneumoniae and Mycoplasma genitalium in the presence of human DNA. J Gen Microbiol 139:2431–2437 [CrossRef]
    [Google Scholar]
  3. Crowcroft N. S., Andrews N., Rooney C., Brisson M., Miller E. 2002; Deaths from pertussis are underestimated in England. Arch Dis Child 86:336–338 [CrossRef]
    [Google Scholar]
  4. Crowcroft N. S., Booy R., Harrison T. & 8 other authors; 2003; Severe and unrecognised: pertussis in UK infants. Arch Dis Child 88:802–806 [CrossRef]
    [Google Scholar]
  5. Douglas E., Coote J. G., Parton R., McPheat W. 1993; Identification of Bordetella pertussis in nasopharyngeal swabs by PCR amplification of a region of the adenylate cyclase gene. J Med Microbiol 38:140–144 [CrossRef]
    [Google Scholar]
  6. Giammanco A., Taormina S., Chiarini A., Dardanoni G., Stefanelli P., Salmaso S., Mastrantonio P. 2003a; Analogous IgG subclass response to pertussis toxin in vaccinated children, healthy or affected by whooping cough. Vaccine 21:1924–1931 [CrossRef]
    [Google Scholar]
  7. Giammanco A., Chiarini A., Maple P. A. C. & 10 other authors (2003b). European Sero-Epidemiology Network: standardisation of the assay results for pertussis. Vaccine 22:112–120 [CrossRef]
    [Google Scholar]
  8. Gladbach S., Hanauer S., Reischl U., Wilson K., Sanden G. 2002; Identification of IS 481 in Bordetella bronchiseptica : implications for Bordetella spp . phylogeny and diagnosis of Bordetella pertussis infections by polymerase chain reaction assays. 7th International Symposium on Pertussis: Genome, Pathogenesis, and Immunity Hinxton, UK:
    [Google Scholar]
  9. Glare E. M., Paton J. C., Premier R. R., Lawrence A. J., Nisbet I. T. 1990; Analysis of a repetitive DNA sequence from Bordetella pertussis and its application to the diagnosis of pertussis using the polymerase chain reaction. J Clin Microbiol 28:1982–1987
    [Google Scholar]
  10. Houard S., Hackel C., Herzog A., Bollen A. 1989; Specific identification of Bordetella pertussis by the polymerase chain reaction. Res Microbiol 140:477–487 [CrossRef]
    [Google Scholar]
  11. Kerr J. R., Matthews R. C. 2000; Bordetella pertussis infection: pathogenesis, diagnosis, management, and the role of protective immunity. Eur J Clin Microbiol Infect Dis 19:77–88 [CrossRef]
    [Google Scholar]
  12. Li Z., Jansen D. L., Finn T. M., Halperin S. A., Kasina A., O'Connor S. P., Aoyama T., Manclark C. R., Brennan M. J. 1994; Identification of Bordetella pertussis infection by shared-primer PCR. J Clin Microbiol 32:783–789
    [Google Scholar]
  13. Mastrantonio P., Stefanelli P., Giuliano M. 1996; Polymerase chain reaction for the detection of Bordetella pertussis in clinical nasopharyngeal aspirates. J Med Microbiol 44:261–266 [CrossRef]
    [Google Scholar]
  14. Meade B. D., Bollen A. 1994; Recommendations for use of the polymerase chain reaction in the diagnosis of Bordetella pertussis infections. J Med Microbiol 41:51–55 [CrossRef]
    [Google Scholar]
  15. Miller E., Fleming D. M., Ashworth L. A. E., Mabbett D. A., Vurdien J. E., Elliot T. S. J. 2000; Serological evidence of pertussis in patients presenting with cough in general practice in Birmingham. Commun Dis Public Health 3:132–134
    [Google Scholar]
  16. Müller F.-M. C., Hoppe J. E., von König C.-H. W. 1997; Laboratory diagnosis of pertussis: state of the art in 1997. J Clin Microbiol 35:2435–2443
    [Google Scholar]
  17. Nardone A., Pebody R. G., Maple P. A. C., Andrews N., Gay N. J., Miller E. 2004; Sero-epidemiology of Bordetella pertussis in England and Wales. Vaccine 22:1314–1319 [CrossRef]
    [Google Scholar]
  18. Parkhill J., Sebaihia M., Preston A. & 50 other authors; 2003; Comparative analysis of the genome sequences of Bordetella pertussis , Bordetella parapertussis and Bordetella bronchiseptica . Nat Genet 35:32–40 [CrossRef]
    [Google Scholar]
  19. Reischl U., Lehn N., Sanden G. N., Loeffelholz M. J. 2001; Real-time PCR assay targeting IS 481 of Bordetella pertussis and molecular basis for detecting Bordetella holmesii . J Clin Microbiol 39:1963–1966 [CrossRef]
    [Google Scholar]
  20. Schmidt-Schläpfer G., Liese J. G., Porter F., Stojanov S., Just M., Belohradsky B. H. 1997; Polymerase chain reaction (PCR) compared with conventional identification in culture for detection of Bordetella pertussis in 7153 children. Clin Microbiol Infect 3:462–467 [CrossRef]
    [Google Scholar]
  21. Tilley P. A. G., Kanchana M. V., Knight I., Blondeau J., Antonishyn N., Deneer H. 2000; Detection of Bordetella pertussis in a clinical laboratory by culture, polymerase chain reaction, and direct fluorescent antibody staining; accuracy, and cost. Diagn Microbiol Infect Dis 37:17–23 [CrossRef]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jmm/10.1099/jmm.0.45624-0
Loading
/content/journal/jmm/10.1099/jmm.0.45624-0
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error