Role of PCR in the diagnosis of pertussis infection in infants: 5 years' experience of provision of a same-day real-time PCR service in England and Wales from 2002 to 2007 Free

Abstract

As part of an enhanced surveillance programme for pertussis in England and Wales, a real-time PCR service for the detection of was introduced for infants aged ≤6 months admitted to a paediatric intensive care unit or paediatric ward with a respiratory illness compatible with pertussis. Two real-time fluorescent resonance energy transfer hybridization probe LightCycler (Roche Diagnostics) PCR assays were used. One (designed in-house) targeted the pertussis toxin S1 promoter (-pr), and included an internal process control to test for sample inhibition and reagent performance. The other (already published) targeted the insertion element IS. The analytical sensitivities of the assays were 100 and 10 fg per reaction for the -pr and IS PCRs, respectively. The -pr assay was specific for , whilst the IS PCR also showed some cross-reactivity with and the type strain of . From April 2002 to March 2007, 848 samples were received from 774 patients and DNA was extracted. Of 824 samples that were suitable for testing, 183 (22.2 %) had evidence of infection (18.9 % -pr and IS; 3.3 % IS only), 621 (75.4 %) were negative and 20 (2.4 %) were inhibitory for the PCR. Within the targeted age group of ≤6 months, most patients (130/138) with evidence of spp. by PCR were ≤3 months old. The overall percentage increase in laboratory-confirmed cases due to PCR compared with culture for the 5 year period described ranged from 9 to 26 % per year (mean 19 %). Real-time PCR is an invaluable tool both for enhanced epidemiological surveillance and for the provision of a rapid diagnosis of pertussis where results can affect patient and contact management.

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2009-08-01
2024-03-29
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References

  1. Alexander E. M., Travis S., Booms C., Kaiser A., Fry N. K., Harrison T. G., Ganpot B., Klein J. L. 2008; Pertussis outbreak on a neonatal unit: identification of a healthcare worker as the likely source. J Hosp Infect 69:131–134 [CrossRef]
    [Google Scholar]
  2. André P., Caro V., Njamkepo E., Wendelboe A. M., Van Rie A., Guiso N. 2008; Comparison of serological and real-time PCR assays to diagnose Bordetella pertussis infection in 2007. J Clin Microbiol 46:1672–1677 [CrossRef]
    [Google Scholar]
  3. Antila M., He Q., de Jong C., Aarts I., Verbakel H., Bruisten S., Keller S., Haanperä M., Mäkinen J. other authors 2006; Bordetella holmesii DNA is not detected in nasopharyngeal swabs from Finnish and Dutch patients with suspected pertussis. J Med Microbiol 55:1043–1051 [CrossRef]
    [Google Scholar]
  4. Bamberger E., Lahat N., Gershtein V., Gershtein R., Benilevi D., Shapiro S., Kassis I., Rubin L., Srugo I. 2005; Diagnosing pertussis: the role of polymerase chain reaction. Isr Med Assoc J 7:351–354
    [Google Scholar]
  5. Crowcroft N. S., Booy R., Harrison T., Spicer L., Britto J., Mok Q., Heath P., Murdoch I., Zambon M. other authors 2003; Severe and unrecognised: pertussis in UK infants. Arch Dis Child 88:802–806 [CrossRef]
    [Google Scholar]
  6. 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]
  7. Dragsted D. M., Dohn B., Madsen J., Jensen J. S. 2004; Comparison of culture and PCR for detection of Bordetella pertussis and Bordetella parapertussis under routine laboratory conditions. J Med Microbiol 53:749–754 [CrossRef]
    [Google Scholar]
  8. Fry N. K., Tzivra O., Li Y. T., McNiff A., Doshi N., Maple P. A. C., Crowcroft N. S., Miller E., George R. C., Harrison T. G. 2004; Laboratory diagnosis of pertussis infections: the role of PCR and serology. J Med Microbiol 53:519–525 [CrossRef]
    [Google Scholar]
  9. 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]
  10. Giammanco A., Chiarini A., Maple P. A. C., Andrews N., Pebody R., Gay N., Ölander R. M., Fivet-Groyne F., Baron S. other authors 2003b; European Sero-Epidemiology Network: standardisation of the assay results for pertussis. Vaccine 22:112–120 [CrossRef]
    [Google Scholar]
  11. 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]
  12. Grimprel E., Bégué P., Anjak I., Betsou F., Guiso N. 1993; Comparison of polymerase chain reaction, culture, and Western immunoblot serology for diagnosis of Bordetella pertussis infection. J Clin Microbiol 31:2745–2750
    [Google Scholar]
  13. Harnden A., Grant C., Harrison T., Perera R., Brueggemann A. B., Mayon-White R., Mant D. 2006; Whooping cough in school age children with persistent cough: prospective cohort study in primary care. BMJ 333:174–177 [CrossRef]
    [Google Scholar]
  14. He Q., Mertsola J., Soini H., Skurnik M., Ruuskanen O., Viljanen M. K. 1993; Comparison of polymerase chain reaction with culture and enzyme immunoassay for diagnosis of pertussis. J Clin Microbiol 31:642–645
    [Google Scholar]
  15. Herwaldt L. A. 1991; Pertussis in adults. What physicians need to know. Arch Intern Med 151:1510–1512 [CrossRef]
    [Google Scholar]
  16. 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]
  17. HPA 2007 Health Protection Report 2, 9–10 London: Health Protection Agency; http://www.hpa.org.uk/hpr/archives/2008/hpr4808.pdf
    [Google Scholar]
  18. HPA 2008 Statutory Notifications of Infectious Diseases (NOIDs) – Annual Totals 1994 to 2007 – England and Wales London: Health Protection Agency; http://www.hpa.org.uk/web/HPAweb&HPAwebStandard/HPAweb_C/1195733756346
    [Google Scholar]
  19. 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]
  20. Kösters K., Reischl U., Schmetz J., Riffelmann M., Von Konig C. H. W. 2002; Real-time LightCycler PCR for detection and discrimination of Bordetella pertussis and Bordetella parapertussis . J Clin Microbiol 40:1719–1722 [CrossRef]
    [Google Scholar]
  21. 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]
  22. Litt D. J., Samuel D., Duncan J., Harnden A., George R. C., Harrison T. G. 2006; Detection of anti-pertussis toxin IgG in oral fluids for use in diagnosis and surveillance of Bordetella pertussis infection in children and young adults. J Med Microbiol 55:1223–1228 [CrossRef]
    [Google Scholar]
  23. Litt D. J., Neal S. E., Fry N. K. 2009; Changes in genetic diversity of the Bordetella pertussis population in the United Kingdom between 1920 and 2006 reflect vaccination coverage and the emergence of a single dominant clonal type. J Clin Microbiol 47:680–688 [CrossRef]
    [Google Scholar]
  24. 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]
  25. Matthews R. 1997; The diagnosis of pertussis infections: a recurring challenge. Publ Health Lab Serv Microbiol Digest 14:79–84
    [Google Scholar]
  26. Muyldermans G., Soetens O., Antoine M., Bruisten S., Vincart B., Doucet-Populaire F., Fry N. K., Olcén P., Scheftel J. M. other authors 2005; External quality assessment for molecular detection of Bordetella pertussis in European laboratories. J Clin Microbiol 43:30–35 [CrossRef]
    [Google Scholar]
  27. Nelson J. D. 1978; The changing epidemiology of pertussis in young infants. The role of adults as reservoirs of infection. Am J Dis Child 132:371–373 [CrossRef]
    [Google Scholar]
  28. Packard E. R., Parton R., Coote J. G., Fry N. K. 2004; Sequence variation and conservation in virulence-related genes of Bordetella pertussis isolates from the UK. J Med Microbiol 53:355–365 [CrossRef]
    [Google Scholar]
  29. Probert W. S., Ely J., Schrader K., Atwell J., Nossoff A., Kwan S. 2008; Identification and evaluation of new target sequences for specific detection of Bordetella pertussis by real-time PCR. J Clin Microbiol 46:3228–3231 [CrossRef]
    [Google Scholar]
  30. Reischl U., Burggraf S., Leppmeier B., Linde H.-J., Lehn N. 2001a; Rapid and specific detection of Bordetella pertussis in clinical specimens by LightCycler PCR. In Rapid Cycle Real-Time PCR: Methods and Applications pp 313–321 Edited by Meuer S., Wittwer C., Nakagawara K.-I. Berlin: Springer;
    [Google Scholar]
  31. Reischl U., Lehn N., Sanden G. N., Loeffelholz M. J. 2001b; 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]
  32. Sambrook J., Russell D. W. 2001; Molecular Cloning: a Laboratory Manual . , 3rd edn. A8.12–14 Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
  33. Senanayake S. 2007; Pertussis in Australia today – a disease of adolescents and adults that can kill infants. Aust Fam Physician 36:51–56
    [Google Scholar]
  34. Sheppard C. L., Harrison T. G., Morris R., Hogan A., George R. C. 2004; Autolysin-targeted LightCycler assay including internal process control for detection of Streptococcus pneumoniae DNA in clinical samples. J Med Microbiol 53:189–195 [CrossRef]
    [Google Scholar]
  35. Van Buynder P. G., Owen D., Vurdien J. E., Andrews N. J., Matthews R. C., Miller E. 1999; Bordetella pertussis surveillance in England and Wales: 1995–7. Epidemiol Infect 123:403–411 [CrossRef]
    [Google Scholar]
  36. Von Wintzingerode F., Schattke A., Siddiqui R. A., Rösick U., Göbel U. B., Gross R. 2001; Bordetella petrii sp. nov., isolated from an anaerobic bioreactor, and emended description of the genus Bordetella . Int J Syst Evol Microbiol 51:1257–1265
    [Google Scholar]
  37. Ward J. I., Cherry J. D., Chang S.-J., Partridge S., Lee H., Treanor J., Greenberg D. P., Keitel W., Barenkamp S. other authors 2005; Efficacy of an acellular pertussis vaccine among adolescents and adults. N Engl J Med 353:1555–1563 [CrossRef]
    [Google Scholar]
  38. WHO 2005; Pertussis vaccines – WHO position paper. Wkly Epidemiol Rec 80:31–39
    [Google Scholar]
  39. Yih W. K., Silva E. A., Ida J., Harrington N., Lett S. M., George H. 1999; Bordetella holmesii -like organisms isolated from Massachusetts patients with pertussis-like symptoms. Emerg Infect Dis 5:441–443 [CrossRef]
    [Google Scholar]
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