Pesquisa do antígeno circulante de Cryptococcus neoformans em líquido cefalorraqueano pelo teste de coaglutinação

Cano, Maria Isabel Nogueira; Livramento, José Antonio

doi:10.1590/S0036-46651990000600011

Resumos

Foram utilizadas 82 LCR de transplantados renais (24 pacientes), 43 LCR de pacientes com críptococose comprovada (controles positivos), 35 LCR de pacientes com outras doenças (histoplasmose, paracoccidioidomicose e infecções bacterianas) como controles negativos. Os primeiros foram cultivados em ágar Sabouraud com sementes de girassol e juntamente com os demais examinado pelo teste de látex para pesquisa de antígeno circulante de C. neoformans, qualitativamente. O teste de Coaglutinação foi realizado qualitativamente e quantitativamente, encontrando-se títulos até a diluição 1:2048. Não foram detectadas reações falso-positivas ou falso-negativas entre os controles. Como prova de valor diagnóstico demonstrou: sensibilidade - 92,1%; especificidade - 92,6% e eficiência - 92,3%. Provou também ser um teste rápido, exato e econômico, embora sua escolha dependa do pré-tratamento de LCR (80ºC por 3 a 5 minutos) e soros (diluição ou álcali-precipitação) para evitar autoaglutinação e aumentar a sensibilidade da reação.

We tested 82 CSF of 24 renal transplanted patients (RT) with cerebral cryptococcosis, 8 CSF of asymptomatic RT patients, 43 CSF of proven cryptococcosis cases (positive control) and 35 CSF of patients with other diseases (histoplasmosis, paracoccidioidomycosis and bacterial infections) as negative control. The RT CSF were cultured in Sabouraud agar slant added with sunflower seeds and both control and RT CSF were qualitatively examined by cryptococcosis latex test (Crypto-LA test). The COA test was developed both qualitatively and quantitatively. The highest titre encountered was 1:2048. No false reactions appeared among the controls. The diagnostic value demonstrated by Galen and Gambino's method was: sensitivity - 92.1%; specificity - 92.6% and efficiency - 92.3%. Besides that, the COA proved to be quick, exact and cheap, but it depends on CSF and sera pre-treatment, in order to avoid autoagglutination and increase its sensibility.

Coagglutination Test; Cryptococcus neoformans; Antigen detection

SEROLOGY

Coaglutination test (COA) for Cryptococcus neoformans circulating antigen detection in cerebral spinal fluid (CSF)

Pesquisa do antígeno circulante de Cryptococcus neoformans em líquido cefalorraqueano pelo teste de coaglutinação

Maria Isabel Nogueira Cano^I; José Antonio Livramento^II

^ILaboratório de Micologia Médica do Instituto de Medicina Tropical de São Paulo e LIM/53 HC-FMUSP. São Paulo, SP, Brasil

^IILaboratório de Investigação em Neurologia, LIM/15 HC-FMUSP. São Paulo, SP, Brasil

^{Address for correspondence} Address for correspondence: Maria Isabel Nogueira CANO Faculdade de Medicina da Universidade de São Paulo, Laboratório de Micologia Médica Av. Dr. Arnaldo, 455 CEP 01246 São Paulo, SP, Brasil

SUMMARY

We tested 82 CSF of 24 renal transplanted patients (RT) with cerebral cryptococcosis, 8 CSF of asymptomatic RT patients, 43 CSF of proven cryptococcosis cases (positive control) and 35 CSF of patients with other diseases (histoplasmosis, paracoccidioidomycosis and bacterial infections) as negative control.

The RT CSF were cultured in Sabouraud agar slant added with sunflower seeds and both control and RT CSF were qualitatively examined by cryptococcosis latex test (Crypto-LA test).

The COA test was developed both qualitatively and quantitatively. The highest titre encountered was 1:2048. No false reactions appeared among the controls. The diagnostic value demonstrated by Galen and Gambino's method was: sensitivity - 92.1%; specificity - 92.6% and efficiency - 92.3%. Besides that, the COA proved to be quick, exact and cheap, but it depends on CSF and sera pre-treatment, in order to avoid autoagglutination and increase its sensibility.

Key words: Coagglutination Test; Cryptococcus neoformans; Antigen detection.

RESUMO

Foram utilizadas 82 LCR de transplantados renais (24 pacientes), 43 LCR de pacientes com críptococose comprovada (controles positivos), 35 LCR de pacientes com outras doenças (histoplasmose, paracoccidioidomicose e infecções bacterianas) como controles negativos. Os primeiros foram cultivados em ágar Sabouraud com sementes de girassol e juntamente com os demais examinado pelo teste de látex para pesquisa de antígeno circulante de C. neoformans, qualitativamente.

O teste de Coaglutinação foi realizado qualitativamente e quantitativamente, encontrando-se títulos até a diluição 1:2048. Não foram detectadas reações falso-positivas ou falso-negativas entre os controles. Como prova de valor diagnóstico demonstrou: sensibilidade - 92,1%; especificidade - 92,6% e eficiência - 92,3%. Provou também ser um teste rápido, exato e econômico, embora sua escolha dependa do pré-tratamento de LCR (80ºC por 3 a 5 minutos) e soros (diluição ou álcali-precipitação) para evitar autoaglutinação e aumentar a sensibilidade da reação.

Full text available only in PDF format.

Texto completo disponível apenas em PDF.

ACKNOWLEDGEMENTS

We gratefully acknowledge Dr. Gildo Del Negro and Dr. Maria Conceição Rodrigues for correcting the manuscript.

Recebido para publicação em 21/5/1990.

1. BAVA, J. Coaglutinación para la determinación de antigeno capsular de Cryptococcus neoformans en fluidos biológicos. Rev. argent. Micol., 9: 33-34, 1986.
2. BLOOMFIELD, N.; GORDON, M.A. & ELMENDORF, F. Detection for Cryptococcus neoformans antigen in body fluids by latex agglutination. Proc. Soc. exp. Biol. (N. Y.), 144: 64-67, 1963.
3. CHRISTENSEN, P.; KHALMEHER, O.; JONSSON, S. & KRONVALL, O. New method for serological grouping of streptococci with specific antibodies adsorbed protein A containing Staphilococci. Infect. Immun., 7: 881-885, 1973.
4. CIVILA, E. & DÍAZ, I.A. Aislamiento de Cryptococcus neoformans de excretas de palomas en la ciudad de Montevideo. Rev. urug. Pat. clin. Microbiol., 14: 41-48, 1976.
5. EDWARDS, E.A. & LARSON, G.L. New method of grouping beta hemolitic Streptococci directly on sheep blood agar plate by coagglutination of specifically sensitized protein A containing Staphilococci. Appl. Microbiol., 28: 972-976, 1974.
6. GALEN, R.S. & GAMBINO, S.R. Beyond normality: the predictive value and efficiency of medical diagnosis. New York, Wiley Biomedical Publication, 1975.
7. GOLDSCHMIDT, M.C. Use of coagglutination technics in the rapid identification of microorganisms. In: GOLDSCHMIDT, M.C. Rapid methods and automation in microbiology. Washington, American Society for Microbiology; R. C. Tilton, 1982. p. 202-206.
8. KAUFMAN, L. & BLUMER, S. Cryptococcosis: the awakening giant. In: The Black and White yeasts. Proceedings of the Fourth International Conference on the Mycosis. Washington. Pan American Health Organization, 1978. p. 176-182.
9. KAUFMAN, L. Mycoserology: Its vital role in diagnosing systemic mycotic infections. Jap. J. med. Mycol.. 24: 1-8, 1983.
10. KOSHI, G.; THANGAVELU, C.P. & BRAHMADATHAN, K.N. The reliability and rapidity of the coagglutination technic and its comparison with precipitin technics in the group of Streptococci. Amer. J. clin. Path., 71: 709-712, 1977.
11. KOSHI, G.; ANANDI, V.; MAMMEN, C. & PUNTHEN-VEETTIL, S.J. Development of a coagglutination (COA) technic to detect Candida antigenemia. Amer. J. clin. Path., 88: 424-435, 1987.
12. LOWRY, O.H.; ROSENBROUWGH, M.J.; FARR, A.L. & RANDALL, R.J. Protein measurement with the folin phenol reagent. J. biol. Chem., 193: 279-281, 1980.
13. MACKENZIE, D.W.R. Serodiagnosis. In: HOWARD, D.H. & HOWARD, L.E. Fungi pathogenic for humans and animals. Part B Pathogenicity and detection. New York, Marcel Dekker, 1983. p. 164-169.
14. MASIH, D.; NEGRONI, R.; NOBILE, R.; RIERA, C. & RUBSTEIN, H. Nueva criptococcina para intradermorreacciones. Rev. argent. Micol., 6: 19-23, 1983.
15. PALMER, D.F.; KAUFMAN, L.; KAPLAN, W. & CAVALLARO, J.J. Slide latex agglutination test for cryptococcal antigen. In: PALMER, D.F.; KAUFMAN, L.; KAPLAN, W. & CAVALLARO, J.J. Serodiagnosis of mycotic disease. Springfield, Charles C. Thomas, 1977. p. 94-106.
16. SCOTT, E.N.; MUCHMORE, H.G. & FELTON, F.G. Comparison of enzyme immunoassay and latex agglutination methods for detection of Cryptococcus neoformans antigen. Amer. J. clin. Path., 73: 790-794, 1980.

Address for correspondence:

Maria Isabel Nogueira CANO

Faculdade de Medicina da Universidade de São Paulo, Laboratório de Micologia Médica

Av. Dr. Arnaldo, 455

CEP 01246 São Paulo, SP, Brasil

Datas de Publicação

Publicação nesta coleção
11 Set 2006
Data do Fascículo
Dez 1990

Histórico

Recebido
21 Maio 1990

This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.

[1] 1. BAVA, J. Coaglutinación para la determinación de antigeno capsular de Cryptococcus neoformans en fluidos biológicos. Rev. argent. Micol., 9: 33-34, 1986.

[2] 2. BLOOMFIELD, N.; GORDON, M.A. & ELMENDORF, F. Detection for Cryptococcus neoformans antigen in body fluids by latex agglutination. Proc. Soc. exp. Biol. (N. Y.), 144: 64-67, 1963.

[3] 3. CHRISTENSEN, P.; KHALMEHER, O.; JONSSON, S. & KRONVALL, O. New method for serological grouping of streptococci with specific antibodies adsorbed protein A containing Staphilococci. Infect. Immun., 7: 881-885, 1973.

[4] 4. CIVILA, E. & DÍAZ, I.A. Aislamiento de Cryptococcus neoformans de excretas de palomas en la ciudad de Montevideo. Rev. urug. Pat. clin. Microbiol., 14: 41-48, 1976.

[5] 5. EDWARDS, E.A. & LARSON, G.L. New method of grouping beta hemolitic Streptococci directly on sheep blood agar plate by coagglutination of specifically sensitized protein A containing Staphilococci. Appl. Microbiol., 28: 972-976, 1974.

[6] 6. GALEN, R.S. & GAMBINO, S.R. Beyond normality: the predictive value and efficiency of medical diagnosis. New York, Wiley Biomedical Publication, 1975.

[7] 7. GOLDSCHMIDT, M.C. Use of coagglutination technics in the rapid identification of microorganisms. In: GOLDSCHMIDT, M.C. Rapid methods and automation in microbiology. Washington, American Society for Microbiology; R. C. Tilton, 1982. p. 202-206.

[8] 8. KAUFMAN, L. & BLUMER, S. Cryptococcosis: the awakening giant. In: The Black and White yeasts. Proceedings of the Fourth International Conference on the Mycosis. Washington. Pan American Health Organization, 1978. p. 176-182.

[9] 9. KAUFMAN, L. Mycoserology: Its vital role in diagnosing systemic mycotic infections. Jap. J. med. Mycol.. 24: 1-8, 1983.

[10] 10. KOSHI, G.; THANGAVELU, C.P. & BRAHMADATHAN, K.N. The reliability and rapidity of the coagglutination technic and its comparison with precipitin technics in the group of Streptococci. Amer. J. clin. Path., 71: 709-712, 1977.

[11] 11. KOSHI, G.; ANANDI, V.; MAMMEN, C. & PUNTHEN-VEETTIL, S.J. Development of a coagglutination (COA) technic to detect Candida antigenemia. Amer. J. clin. Path., 88: 424-435, 1987.

[12] 12. LOWRY, O.H.; ROSENBROUWGH, M.J.; FARR, A.L. & RANDALL, R.J. Protein measurement with the folin phenol reagent. J. biol. Chem., 193: 279-281, 1980.

[13] 13. MACKENZIE, D.W.R. Serodiagnosis. In: HOWARD, D.H. & HOWARD, L.E. Fungi pathogenic for humans and animals. Part B Pathogenicity and detection. New York, Marcel Dekker, 1983. p. 164-169.

[14] 14. MASIH, D.; NEGRONI, R.; NOBILE, R.; RIERA, C. & RUBSTEIN, H. Nueva criptococcina para intradermorreacciones. Rev. argent. Micol., 6: 19-23, 1983.

[15] 15. PALMER, D.F.; KAUFMAN, L.; KAPLAN, W. & CAVALLARO, J.J. Slide latex agglutination test for cryptococcal antigen. In: PALMER, D.F.; KAUFMAN, L.; KAPLAN, W. & CAVALLARO, J.J. Serodiagnosis of mycotic disease. Springfield, Charles C. Thomas, 1977. p. 94-106.

[16] 16. SCOTT, E.N.; MUCHMORE, H.G. & FELTON, F.G. Comparison of enzyme immunoassay and latex agglutination methods for detection of Cryptococcus neoformans antigen. Amer. J. clin. Path., 73: 790-794, 1980.

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