Skip to main content
PMC home page
Journal of Clinical Microbiology logoLink to Journal of Clinical Microbiology
. 1997 Dec;35(12):3215–3219. doi: 10.1128/jcm.35.12.3215-3219.1997

Confirmation of suspicious cases of meningococcal meningitis by PCR and enzyme-linked immunosorbent assay.

N B Saunders 1, D R Shoemaker 1, B L Brandt 1, W D Zollinger 1
PMCID: PMC230150  PMID: 9399522

Abstract

A significant problem in efficacy trials of meningococcal vaccines has been accurate identification of all cases of meningococcal disease that occur in study populations. The accuracy of case determination would be improved by utilizing methods which confirm or disprove suspicious cases of meningococcal disease that are culture negative. A collection of serum and cerebrospinal fluid (CSF) samples from a meningococcal vaccine field trial performed in Iquique, Chile, were utilized to assess the status of patients for whom cultures, Gram stains, and clinical evaluations for meningococcal disease were available. Nested PCRs (nPCRs) for amplification of Neisseria meningitidis DNA in CSF samples and enzyme-linked immunosorbent assays (ELISAs) for quantification of serum immunoglobulin G antibodies specific for N. meningitidis were used in combination to confirm or eliminate cases classified by physicians as suspicious for meningococcal disease. Samples from 12 of 79 patients suspected of having meningococcal meningitis tested positive by both methods; specimens from 61 of the 79 were negative by both methods; and samples from 6 patients yielded ambiguous results, and these cases remained unconfirmed. Direct sequence analysis of amplified DNA from patients suspected of having meningococcal disease confirmed that 2 of the 12 newly confirmed cases were not attributable to the typical epidemic strain (B:15:P1.[7],3) while the others were due to the epidemic strain. A combination of nPCR and ELISA reduced the number of suspicious cases in this study from 79 to 6, thereby improving the potential for assessment of vaccine efficacy. Molecular identification by nPCR in conjunction with immunological assessment of patient response could be considered diagnostic of disease in future testing of meningococcal vaccines to improve efficacy analyses.

Full Text

The Full Text of this article is available as a PDF (143.6 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Boslego J., Garcia J., Cruz C., Zollinger W., Brandt B., Ruiz S., Martinez M., Arthur J., Underwood P., Silva W. Efficacy, safety, and immunogenicity of a meningococcal group B (15:P1.3) outer membrane protein vaccine in Iquique, Chile. Chilean National Committee for Meningococcal Disease. Vaccine. 1995 Jun;13(9):821–829. doi: 10.1016/0264-410x(94)00037-n. [DOI] [PubMed] [Google Scholar]
  2. Cartwright K., Reilly S., White D., Stuart J. Early treatment with parenteral penicillin in meningococcal disease. BMJ. 1992 Jul 18;305(6846):143–147. doi: 10.1136/bmj.305.6846.143. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Caugant D. A., Høiby E. A., Frøholm L. O., Brandtzaeg P. Polymerase chain reaction for case ascertainment of meningococcal meningitis: application to the cerebrospinal fluids collected in the course of the Norwegian meningococcal serogroup B protection trial. Scand J Infect Dis. 1996;28(2):149–153. doi: 10.3109/00365549609049066. [DOI] [PubMed] [Google Scholar]
  4. Cruz C., Pavez G., Aguilar E., Grawe L., Cam J., Mendez F., Garcia J., Ruiz S., Vicent P., Canepa I. Serotype-specific outbreak of group B meningococcal disease in Iquique, Chile. Epidemiol Infect. 1990 Aug;105(1):119–126. doi: 10.1017/s0950268800047713. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Gray L. D., Fedorko D. P. Laboratory diagnosis of bacterial meningitis. Clin Microbiol Rev. 1992 Apr;5(2):130–145. doi: 10.1128/cmr.5.2.130. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Kwok S., Higuchi R. Avoiding false positives with PCR. Nature. 1989 May 18;339(6221):237–238. doi: 10.1038/339237a0. [DOI] [PubMed] [Google Scholar]
  7. McGuinness B. T., Clarke I. N., Lambden P. R., Barlow A. K., Poolman J. T., Jones D. M., Heckels J. E. Point mutation in meningococcal por A gene associated with increased endemic disease. Lancet. 1991 Mar 2;337(8740):514–517. doi: 10.1016/0140-6736(91)91297-8. [DOI] [PubMed] [Google Scholar]
  8. Rao V. B., Saunders N. B. A rapid polymerase-chain-reaction-directed sequencing strategy using a thermostable DNA polymerase from Thermus flavus. Gene. 1992 Apr 1;113(1):17–23. doi: 10.1016/0378-1119(92)90665-c. [DOI] [PubMed] [Google Scholar]
  9. Rosenqvist E., Høiby E. A., Wedege E., Caugant D. A., Frøholm L. O., McGuinness B. T., Brooks J., Lambden P. R., Heckels J. E. A new variant of serosubtype P1.16 in Neisseria meningitidis from Norway, associated with increased resistance to bactericidal antibodies induced by a serogroup B outer membrane protein vaccine. Microb Pathog. 1993 Sep;15(3):197–205. doi: 10.1006/mpat.1993.1070. [DOI] [PubMed] [Google Scholar]
  10. Saunders N. B., Zollinger W. D., Rao V. B. A rapid and sensitive PCR strategy employed for amplification and sequencing of porA from a single colony-forming unit of Neisseria meningitidis. Gene. 1993 Dec 31;137(2):153–162. doi: 10.1016/0378-1119(93)90001-j. [DOI] [PubMed] [Google Scholar]
  11. Sjursen H., Wedege E., Rosenqvist E., Naess A., Halstensen A., Matre R., Solberg C. O. IgG subclass antibodies to serogroup B meningococcal outer membrane antigens following infection and vaccination. APMIS. 1990 Dec;98(12):1061–1069. doi: 10.1111/j.1699-0463.1990.tb05035.x. [DOI] [PubMed] [Google Scholar]
  12. Sørensen H. T., Møller-Petersen J., Krarup H. B., Pedersen H., Hansen H., Hamburger H. Diagnostic problems with meningococcal disease in general practice. J Clin Epidemiol. 1992 Nov;45(11):1289–1293. doi: 10.1016/0895-4356(92)90169-n. [DOI] [PubMed] [Google Scholar]
  13. Wedege E., Dalseg R., Caugant D. A., Poolman J. T., Frøholm L. O. Expression of an inaccessible P1.7 subtype epitope on meningococcal class 1 proteins. J Med Microbiol. 1993 Jan;38(1):23–28. doi: 10.1099/00222615-38-1-23. [DOI] [PubMed] [Google Scholar]
  14. Wiedbrauk D. L., Werner J. C., Drevon A. M. Inhibition of PCR by aqueous and vitreous fluids. J Clin Microbiol. 1995 Oct;33(10):2643–2646. doi: 10.1128/jcm.33.10.2643-2646.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Zollinger W. D., Mandrell R. E., Griffiss J. M., Altieri P., Berman S. Complex of meningococcal group B polysaccharide and type 2 outer membrane protein immunogenic in man. J Clin Invest. 1979 May;63(5):836–848. doi: 10.1172/JCI109383. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Clinical Microbiology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES