Scandinavian Journal of Infectious Diseases

ISSN: 0036-5548 (Print) 1651-1980 (Online) Journal homepage: http://www.tandfonline.com/loi/infd19

Failure of Diagnosing Group B Meningococcal Meningitis by Immunoelectroosmophoresis Jean-Pierre Ghanassia, Abel Slim, Eugéanie Bergogne-Berezin & Jacques Modai To cite this article: Jean-Pierre Ghanassia, Abel Slim, Eugéanie Bergogne-Berezin & Jacques Modai (1977) Failure of Diagnosing Group B Meningococcal Meningitis by Immunoelectroosmophoresis, Scandinavian Journal of Infectious Diseases, 9:4, 313-314, DOI: 10.3109/inf.1977.9.issue-4.12 To link to this article: http://dx.doi.org/10.3109/inf.1977.9.issue-4.12

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Date: 13 April 2016, At: 20:44

Scand J Infect Dis 9: 313-314, 1977

Short Communication

Failure of Diagnosing Group B Meningococcal Meningitis by ImmuFoelectroosmoplaoresis JEAN-PIERRE GHANASSIA, ABEL SLIM, E U G E N I E BERGOGNE-BEREZIN and JACQUES MODAI

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From the Department of Infectious Diseases, Claude Bernard Hospital, and the Central Microbiological Laboratory, Bichat Hospital, Paris, France

ABSTRACT. The immunoelectroosmophoresis (IEOP) technique has been used to show the presence of bacterial antigens in the cerebrospinal fluid (CSF) of 28 cases of group B meningococcal meningitis. No specific precipitation was observed with any of the 28 CSF examined. Up until now this method has been of no value in the diagnosis of group B meningococcal meningitis.

INTRODUCTION In cases of bacterial meningitis it is important from the standpoints of prognosis, therapy and epidemiology to detect the responsible germ. When isolation is unsuccessful for various reasons (8), it is possible to show the presence of specific antigens in the patient’s cerebrospinal fluid (CSF). This can be achieved by the immunoelectroosmophoresis (IEOP) technique (2, 3, 9, lo), which simply and rapidly gives valuable information within a n hour concerning parasitic, fungal, viral, streptococcal and other bacterial antigens (1, 4, 5, 6). Our particular purpose was t o assess the value of the I EOP method for diagnosing group B meningococcal meningitis since this germ is most often responsible for bacterial meningitis in France.

M A T ER I A L A N D M E T HODS Patients 28 samples of CSF were obtained by lumbar puncture from 28 patients hospitalised in the “Clinique des Maladies Infectieuses” from September 1975 to September 1976. All patients demonstrated clinical evidence of a febrile meningitis syndrome, resulting in need for lumbar puncture. All CSF samples were examined between 6 and 72 h after the onset of the disease. Since we intended to assess the use of IEOP for the diagnosis of group B meningococcal meningitis, we systematically discarded both patients who had been given antibiotics before admission and patients in whom another germ could

be isolated by usual bacteriological techniques (these same samples have been kept for another study). IEOP technique The IEOP method consists of simultaneous electrophoretic migration in one single buffered and gelated medium of both a known antigenic suspension and the CSF sample to be studied. Their reciprocal specificity materializes into a precipitation line in the intermediate region. The antibody used was a meningococcal antiserum of the A and B groups (Wellcome Laboratories). The gel medium used was a 1% agarose gel in buffer solution laid on glass plates. Some experiments were also carried out on acetate cellulose strips. Several buffer solutions were used during this study, in particular sodium veronal buffer, pH 8.6, ionic strength 0.10 and 0.025, and sodium barbital acetate, pH 5.8, ionic strength 0.05 and 0.025. The tank was connected to a source of continuous current (5 and 15 volts per cm). The duration of electrodiffusion was usually 45 min. Both shorter runs (20 and 30 min) and longer runs (60and 90 min) were also carried out. All plates were then placed in water-saturated chambers at 20°C for 12-72 h and were frequently examined for any possible modification. The preparation was washed 3 times with saline solution for 2 h each time, and once with distilled water for 30 min to retain only the insoluble immune complexes. The plates were then dried and stained with amino schwartz.

RESULTS The 28 strains of Neisseria meningitidis isolated were all of the B type. No specific precipitation line was obtained from any of the 28 CSF tested, even though some parts of the experiment were modified concerning duration of assays, variation in voltage, Scand J Infect Dis 9

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J.-P. Ghanassia et al.

variation of pH, changing buffer in the tank, and variation in the position of the CSF and the antiserum.

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DISCUSSION The IEOP technique has a high specificity (5, 9, 10, 1 I). Because no antigen-antibody reaction was detected in our investigation, we also tested the 28 CSF samples with group B meningococcus by Ouchterlony's method, since it may be considered as a reference method which is based on diffusion of samples in a gel medium. It produced the same negative results. The immune sera used should have been able to give positive results since we have observed a precipitate reaction against antigenic extracts of group B meningococcus up to a 1 : 20 dilution. It is well known that the polysaccharide antigens of type B N. meningitidis are only weakly antigenic. We attempted to set free the antigenic fractions in the CSF through ultrasonic treatment of the samples but the results in IEOP remained negative. However, the technique might be improved by concentration of the CSF by a micromethod (7) which might produce a sufficient quantity of group B polysaccharide antigen to give a precipitate, or by microagglutination techniques, which permit detection of much smaller quantities of antigen. It is concluded that classical bacteriological tests are of much greater value to the clinician in the diagnosis of group B meningococcal meningitis than is the IEOP method. A satisfactory IEOP technique would be desirable, because N. meningitidis group B still causes nearly 80% of purulent meningitis in France. ACKNOWLEDGEMENT We are indebted to Dr Fransoise Trtmolitres for revising the manuscript.

REFERENCES I . Bergogne-Berezin, E. & Slim, A. : Le diagnostic de groupe des streptocoques. Ann Biol Clin 34:415, 1976. 2. Bradshaw, W., Schneerson, R., Parke, J. C., Jr & Robbins, J. B.: Bacterial antigen cross-reactive with the capsular polysaccharide of Haemophilus influenzae type b. Lancet 1: 1095, 1971. 3. Fossieck, B., Jr, Craig, R. & Paterson, P. Y.: Counterimmunoelectrophoresis for rapid diagnosis of meningitis due to Diplococcus pneumoniae. J Infect Dis 127: 106, 1973. Scand J Infect Dis 9

4. Greenwood, B. M., Whittle, H. C. & Dominic-Rajkovic, 0.: Countercurrent imrnunoelectrophoresis in the diagnosis of meningococcal infections. Lancet 2:519, 1971. 5. Higashi, G. I . , Sippel, J. E., Girgis, N. I. & Hassan, A.: Counterimmmunoelectrophoresis: An adjunct to bacterial culture in the diagnosis of meningococcal meningitis. Scand J Infect Dis 6: 233, 1974. 6. Jones, D. M. & Abbott, J. D.: Diagnosis of pyogenic meningitis. Lancet 2: 257, 1976. 7. Kahn, S. N. & Thompson, E. J.: New ultramicromethod for concentration of cerebrospinal fluid. Lancet 1: 1275, 1976. 8. Mandal, B. K.: The dilemma of partially treated bacterial meningitis. Scand J Infect Dis 8: 185, 1976. 9. Myhre, E. B.: Rapid diagnosis of bacterial meningitis. Demonstration of bacterial antigen by counterimmunoelectrophoresis. Scand J Infect Dis 6: 237, 1974. 10. Sillanpi%, M., Vaha-Eskeli, E. & Willman, K.: Immunoelectroosrnophoresis (IEOP) for detection of bacterial antigens in cerebrospinal fluid. Scand J Infect Dis 7: 113, 1975. 11. Whittle, H. C., Greenwood, B. M., Davidson, N. MacD., Tomkins, A., Tugwell, P., Warrell, D. A., Zalin, A., Bryceson, A. D. M., Parry, E. H. O., Brueton, M., Duggan, M., Oomen, J. M. V. & Dominic-Rajkovic, 0.: Meningococcal antigen in diagnosis and treatment of group A meningococcal infections. Am J Med 58: 823, 1975. Address f o r reprints: J . Modai, M.D., Clinique des Maladies Infectieuses. H6pital Claude Bernard, 10, Avenue de la Porte d'Aubervilliers, 75019-Paris, France

Failure of diagnosing group B meningococcal meningitis by immunoelectroosmophoresis.

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