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. 1995 Feb;33(2):458–462. doi: 10.1128/jcm.33.2.458-462.1995

Typing Neisseria meningitidis by analysis of restriction fragment length polymorphisms in the gene encoding the class 1 outer membrane protein: application to assessment of epidemics throughout the last 4 decades in China.

P Zhu 1, X Hu 1, L Xu 1
PMCID: PMC227966  PMID: 7714207

Abstract

A typing method was developed for Neisseria meningitidis serogroup A by analysis of restriction fragment length polymorphisms (RFLP) of the class 1 outer membrane protein gene (porA). By using appropriate primers, an approximately 1,116-bp fragment of the porA gene was amplified by PCR and then was digested with the restriction endonuclease MspI. The digestion products were separated on 10% polyacrylamide gels and were stained with silver. One hundred three clinical isolates of group A N. meningitidis from 17 provinces of China collected over a 26-year period were analyzed. Results of MspI-generated RFLP profiles of PCR-amplified porA genes were compared with those obtained by conventional serosubtyping. There was a band of about 400 bp common to all strains examined, and the 103 strains of serogroup A resulted in 22 unique RFLP patterns. The differences in bands could be observed mainly in the range of 120 to 280 bp. The smaller fragments were useful in distinguishing meningococci with the same serosubtype. Three epidemic periods were characterized by the presence of three distinct genotypes (a1, a2, and a3), accounting for 74.5% of the strains examined (3.88, 26.21, and 44.66%, respectively). Three predominant RFLP patterns were correlated epidemiologically with cycles of epidemic meningococcal meningitis and were well-matched to the predominant serosubtypes (P1.9, P1.7, 10, and P1.9) that presented at the same prevalence cycles. The genotyping yielded information that allowed strains from one epidemic to be distinguished from those from another that would have been indistinguishable if only serotyping and serosubtyping were available. Therefore, the PCR-RFLP typing method was very useful in the epidemiologic investigation of group A meningococcal meningitis.

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Selected References

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  1. Barlow A. K., Heckels J. E., Clarke I. N. The class 1 outer membrane protein of Neisseria meningitidis: gene sequence and structural and immunological similarities to gonococcal porins. Mol Microbiol. 1989 Feb;3(2):131–139. doi: 10.1111/j.1365-2958.1989.tb01802.x. [DOI] [PubMed] [Google Scholar]
  2. Beati L., Finidori J. P., Gilot B., Raoult D. Comparison of serologic typing, sodium dodecyl sulfate-polyacrylamide gel electrophoresis protein analysis, and genetic restriction fragment length polymorphism analysis for identification of rickettsiae: characterization of two new rickettsial strains. J Clin Microbiol. 1992 Aug;30(8):1922–1930. doi: 10.1128/jcm.30.8.1922-1930.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bjorvatn B., Lund V., Kristiansen B. E., Korsnes L., Spanne O., Lindqvist B. Applications of restriction endonuclease fingerprinting of chromosomal DNA of Neisseria meningitidis. J Clin Microbiol. 1984 Jun;19(6):763–765. doi: 10.1128/jcm.19.6.763-765.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Caugant D. A., Frøholm L. O., Bøvre K., Holten E., Frasch C. E., Mocca L. F., Zollinger W. D., Selander R. K. Intercontinental spread of a genetically distinctive complex of clones of Neisseria meningitidis causing epidemic disease. Proc Natl Acad Sci U S A. 1986 Jul;83(13):4927–4931. doi: 10.1073/pnas.83.13.4927. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Facinelli B., Biavasco F., Varaldo P. E. Use of DNA fingerprinting in an epidemiologic study of outbreak-specific and non-specific strains of group C Neisseria meningitidis. Eur J Epidemiol. 1990 Mar;6(1):80–83. doi: 10.1007/BF00155555. [DOI] [PubMed] [Google Scholar]
  6. Fox A. J., Jones D. M., Gray S. J., Caugant D. A., Saunders N. A. An epidemiologically valuable typing method for Neisseria meningitidis by analysis of restriction fragment length polymorphisms. J Med Microbiol. 1991 May;34(5):265–270. doi: 10.1099/00222615-34-5-265. [DOI] [PubMed] [Google Scholar]
  7. Frasch C. E., Zollinger W. D., Poolman J. T. Serotype antigens of Neisseria meningitidis and a proposed scheme for designation of serotypes. Rev Infect Dis. 1985 Jul-Aug;7(4):504–510. doi: 10.1093/clinids/7.4.504. [DOI] [PubMed] [Google Scholar]
  8. Frost E. H., Deslandes S., Veilleux S., Bourgaux-Ramoisy D. Typing Chlamydia trachomatis by detection of restriction fragment length polymorphism in the gene encoding the major outer membrane protein. J Infect Dis. 1991 May;163(5):1103–1107. doi: 10.1093/infdis/163.5.1103. [DOI] [PubMed] [Google Scholar]
  9. Frøholm L. O., Bøvre K., Holten E., Zollinger W. D. Serotyping of meningococci by coagglutination with monoclonal antibodies. NIPH Ann. 1983 Dec;6(2):125–131. [PubMed] [Google Scholar]
  10. Hermans P. W., Schuitema A. R., Van Soolingen D., Verstynen C. P., Bik E. M., Thole J. E., Kolk A. H., van Embden J. D. Specific detection of Mycobacterium tuberculosis complex strains by polymerase chain reaction. J Clin Microbiol. 1990 Jun;28(6):1204–1213. doi: 10.1128/jcm.28.6.1204-1213.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Hu X. [Study on periodically prevalent feature for epidemic cerebrospinal meningitis in China]. Zhonghua Liu Xing Bing Xue Za Zhi. 1991 Jun;12(3):136–139. [PubMed] [Google Scholar]
  12. Hu X. [Surveillance of epidemiological features for epidemic cerebrospinal meningitis in China during 1980s]. Zhonghua Liu Xing Bing Xue Za Zhi. 1990 Dec;11(6):321–324. [PubMed] [Google Scholar]
  13. Kertesz D. A., Byrne S. K., Chow A. W. Characterization of Neisseria meningitidis by polymerase chain reaction and restriction endonuclease digestion of the porA gene. J Clin Microbiol. 1993 Oct;31(10):2594–2598. doi: 10.1128/jcm.31.10.2594-2598.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Knight A. I., Cartwright K. A., McFadden J. Identification of a UK outbreak strain of Neisseria meningitidis with a DNA probe. Lancet. 1990 May 19;335(8699):1182–1184. doi: 10.1016/0140-6736(90)92697-g. [DOI] [PubMed] [Google Scholar]
  15. Kristiansen B. E., Ask E., Jenkins A., Fermer C., Rådstrøm P., Skøld O. Rapid diagnosis of meningococcal meningitis by polymerase chain reaction. Lancet. 1991 Jun 29;337(8757):1568–1569. doi: 10.1016/0140-6736(91)93262-8. [DOI] [PubMed] [Google Scholar]
  16. Maiden M. C., Suker J., McKenna A. J., Bygraves J. A., Feavers I. M. Comparison of the class 1 outer membrane proteins of eight serological reference strains of Neisseria meningitidis. Mol Microbiol. 1991 Mar;5(3):727–736. doi: 10.1111/j.1365-2958.1991.tb00743.x. [DOI] [PubMed] [Google Scholar]
  17. McGuinness B. T., Lambden P. R., Heckels J. E. Class 1 outer membrane protein of Neisseria meningitidis: epitope analysis of the antigenic diversity between strains, implications for subtype definition and molecular epidemiology. Mol Microbiol. 1993 Feb;7(4):505–514. doi: 10.1111/j.1365-2958.1993.tb01141.x. [DOI] [PubMed] [Google Scholar]
  18. McGuinness B., Barlow A. K., Clarke I. N., Farley J. E., Anilionis A., Poolman J. T., Heckels J. E. Deduced amino acid sequences of class 1 protein (PorA) from three strains of Neisseria meningitidis. Synthetic peptides define the epitopes responsible for serosubtype specificity. J Exp Med. 1990 Jun 1;171(6):1871–1882. doi: 10.1084/jem.171.6.1871. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Ni H., Knight A. I., Cartwright K., Palmer W. H., McFadden J. Polymerase chain reaction for diagnosis of meningococcal meningitis. Lancet. 1992 Dec 12;340(8833):1432–1434. doi: 10.1016/0140-6736(92)92622-m. [DOI] [PubMed] [Google Scholar]
  20. Niesters H. G., Goessens W. H., Meis J. F., Quint W. G. Rapid, polymerase chain reaction-based identification assays for Candida species. J Clin Microbiol. 1993 Apr;31(4):904–910. doi: 10.1128/jcm.31.4.904-910.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Rodriguez P., Vekris A., de Barbeyrac B., Dutilh B., Bonnet J., Bebear C. Typing of Chlamydia trachomatis by restriction endonuclease analysis of the amplified major outer membrane protein gene. J Clin Microbiol. 1991 Jun;29(6):1132–1136. doi: 10.1128/jcm.29.6.1132-1136.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Saiki R. K., Scharf S., Faloona F., Mullis K. B., Horn G. T., Erlich H. A., Arnheim N. Enzymatic amplification of beta-globin genomic sequences and restriction site analysis for diagnosis of sickle cell anemia. Science. 1985 Dec 20;230(4732):1350–1354. doi: 10.1126/science.2999980. [DOI] [PubMed] [Google Scholar]
  23. Wedege E., Høiby E. A., Rosenqvist E., Frøholm L. O. Serotyping and subtyping of Neisseria meningitidis isolates by co-agglutination, dot-blotting and ELISA. J Med Microbiol. 1990 Mar;31(3):195–201. doi: 10.1099/00222615-31-3-195. [DOI] [PubMed] [Google Scholar]
  24. Williams J. G., Kubelik A. R., Livak K. J., Rafalski J. A., Tingey S. V. DNA polymorphisms amplified by arbitrary primers are useful as genetic markers. Nucleic Acids Res. 1990 Nov 25;18(22):6531–6535. doi: 10.1093/nar/18.22.6531. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Woods C. R., Jr, Versalovic J., Koeuth T., Lupski J. R. Analysis of relationships among isolates of Citrobacter diversus by using DNA fingerprints generated by repetitive sequence-based primers in the polymerase chain reaction. J Clin Microbiol. 1992 Nov;30(11):2921–2929. doi: 10.1128/jcm.30.11.2921-2929.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Zollinger W. D., Moran E. E., Connelly H., Mandrell R. E., Brandt B. Monoclonal antibodies to serotype 2 and serotype 15 outer membrane proteins of Neisseria meningitidis and their use in serotyping. Infect Immun. 1984 Oct;46(1):260–266. doi: 10.1128/iai.46.1.260-266.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. de Marie S., Hoeijmakers J. H., Poolman J. T., Zanen H. C. Filter radioimmunoassay, a method for large-scale serotyping of Neisseria meningitidis. J Clin Microbiol. 1984 Aug;20(2):255–258. doi: 10.1128/jcm.20.2.255-258.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]

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