Evaluation of different methods for Plasmodia detection, in well defined population groups in an endemic area of Brazil

Avila, Sandra L. M.; Leandro, Mirlan C.; Carvalho, Noemia B.; Oliveira, Marilia S.; Arruk, Viviana G.; Sanchez, Maria Carmen A.; Boulos, Marcos; Ferreira, Antonio Walter

doi:10.1590/S0036-46651994000200011

Abstracts

In Brazil, more than 500,000 new cases of malaria were notified in 1992. Plasmodium falciparum and P.vivax are the responsible species for 99.3% of the cases. For adequate treatment, precoce diagnosis is necessary. In this work, we present the results of the traditional Plasmodia detection method, thick blood film (TBF), and the results of alternative methods: Immunofluorescence assay (IFA) with polyclonal antibody and Quantitative Buffy Coat method (QBC)® in a well defined population groups. The analysis were done in relation to the presence or absence of malaria clinical symptoms. Also different classes of immunoglobulins anti-P.falciparum were quantified for the global analysis of the results, mainly in the discrepant results. We concluded that alternative methods are more sensitive than TBF and that the association of epidemiological, clinical and laboratory findings is necessary to define the presence of malaria.

Malaria; Plasmodia detection; diagnosis; IFA; QBC

Mais de 500.000 casos novos de malária foram notificados no Brasil, em 1992. P.falciparum and P.vivax são as espécies responsáveis por 99,3% dos casos. O diagnóstico precoce é indispensável para início do tratamento adequado. Neste trabalho, apresentamos os resultados do método tradicional para detecção de plasmódios, gota espessa, e os resultados de métodos alternativos estudados: imunofluorescência indireta com anticorpo policlonal anti-P.falciparum e QBC-método, em grupos populacionais bem definidos. A análise dos resultados foi feita em relação à presença ou ausênsia de sintomas clínicos de malária. Também, diferentes classes de imunoglobulinas anti-P.falciparum foram quantificadas para auxiliar na análise global dos resultados, principalmente nos resultados discrepantes. Nós concluímos que os métodos alternativos são mais sensíveis do que a gota espessa e que a associação das informações epidemiológicas, clínicas e laboratoriais é necessária para definir a presença de malária.

TECHNICAL TRIALS

Evaluation of different methods for Plasmodia detection, in well defined population groups in an endemic area of Brazil⁽¹ (1 ) The data presented in this work are part of a Ph thesis. ⁾

Avaliação de diferentes métodos para detecção de plasmódios em grupos populacionais bem definidos em uma área endêmica do Brasil

Sandra L. M. Avila^I; Mirlan C. Leandro^I; Noemia B. Carvalho^II; Marilia S. Oliveira^II; Viviana G. Arruk^I; Maria Carmen A. Sanchez^I; Marcos Boulos^I,II; Antonio Walter Ferreira^I,II

^IInstitute of Tropical Medicine of São Paulo - Medical School of the University of São Paulo

^IIInfectious Diseases Department of the Medical School of the University of São Paulo

^{Address for correspondence} Address for correspondence: Sandra do Lago Moraes de Ávila Instituto de Medicina Tropical de São Paulo. Faculdade de Medicina da Universidade de São Paulo Av. Dr. Enéas de Carvalho Aguiar, 470 05403-000 São Paulo, SP, Brasil

SUMMARY

In Brazil, more than 500,000 new cases of malaria were notified in 1992. Plasmodium falciparum and P.vivax are the responsible species for 99.3% of the cases. For adequate treatment, precoce diagnosis is necessary. In this work, we present the results of the traditional Plasmodia detection method, thick blood film (TBF), and the results of alternative methods: Immunofluorescence assay (IFA) with polyclonal antibody and Quantitative Buffy Coat method (QBC)^® in a well defined population groups. The analysis were done in relation to the presence or absence of malaria clinical symptoms. Also different classes of immunoglobulins anti-P.falciparum were quantified for the global analysis of the results, mainly in the discrepant results. We concluded that alternative methods are more sensitive than TBF and that the association of epidemiological, clinical and laboratory findings is necessary to define the presence of malaria.

Keywords: Malaria; Plasmodia detection; diagnosis; IFA; QBC.

RESUMO

Mais de 500.000 casos novos de malária foram notificados no Brasil, em 1992. P.falciparum and P.vivax são as espécies responsáveis por 99,3% dos casos. O diagnóstico precoce é indispensável para início do tratamento adequado. Neste trabalho, apresentamos os resultados do método tradicional para detecção de plasmódios, gota espessa, e os resultados de métodos alternativos estudados: imunofluorescência indireta com anticorpo policlonal anti-P.falciparum e QBC-método, em grupos populacionais bem definidos. A análise dos resultados foi feita em relação à presença ou ausênsia de sintomas clínicos de malária. Também, diferentes classes de imunoglobulinas anti-P.falciparum foram quantificadas para auxiliar na análise global dos resultados, principalmente nos resultados discrepantes. Nós concluímos que os métodos alternativos são mais sensíveis do que a gota espessa e que a associação das informações epidemiológicas, clínicas e laboratoriais é necessária para definir a presença de malária.

Full text available only in PDF format.

Texto completo disponível apenas em PDF.

ACKNOWLEDGEMENTS

This work was in part supported by FAPESP (91/0338-0). We thank to Becton Dickinson for supplying the QBC tubes, Dr. Joseph Perrone for critical review of this manuscript, Mr. Nei Rodrigues de Carvalho Filho for technical assistance and Dr. Fernando Costa Andrade Moraes and Fundação Nacional de Saúde - Regional Mato Grsso.

Recebido para publicação em 29/07/1993.

Aceito para publicação em 19/01/1994.

1. AVILA, S.L.M.; BARNWELL, J.; NUSSENZWEIG, R.S.; ALECRIM, M.G. & FERREIRA, A.W. - Evaluation of the results of the IFI test for Plasmodia detection in patients with malaria. Possibility of application in the blood donors screening. Rev. Soc. bras. Med. trop., 25, (suppl. 2): 64, 1992.
2. AVILA, S.L.M.; MACHADO, M.; LEANDRO, M.C. et al. - Definition of serological profile in the human malaria in the sera of patients with acute and chronic disease by Western Blotting. Rev. Soc. bras. Med. trop., 25, (suppl.. 1): 6, 1992.
3. BARKER JR, R.H.; SUE BSAENG, L.; ROONEY, W. & WIRTH, D.F. - Detection of Plasmodium falciparum infection in human patients: a comparison of the DNA probe method to microscopic diagnosis. Amer. J. trop. Med. Hyg., 41: 266-272, 1989.
4. BASSET, M.T.; TAYLOR, P.; BVIRAKARE, J.; CHITEKA, F. & GOVERE, E. - Clinical diagnosis of malaria: can we improve? J. trop. Med. Hyg., 94: 65-69, 1991.
5. BRUCE-CHWATT, L.J. - From Laveran's discovery to DNA probes: new trends in diagnosis of malaria. Lancet, 2: 1509-1511, 1987.
6. BRUCE-CHWATT, L.J. - Imported malaria: an univited guest. Brit. med. Bull., 38: 179-185, 1982.
7. BRUCE-CHWATT, L.J. - Transfusional malaria revisited. Trop. Dis. Bull., 79: 827-840, 1982.
8. CARVALHO, M.C.R.; SANCHEZ, M.C.A.; QUARTIER, V.P. et al. - Transfusional malaria. Risk of contamination in non endemic areas. Rev. Inst. Med. trop. S. Paulo, 31 (supl. 7): S23, 1989.
9. DUBARRY, M.; LUILIER, M.; MALOT, N. et al. - Enzyme immunoassays for detection of malarial antigens in human plasmas by Plasmodium falciparum monoclonal antibodies. Amer. J. trop. Med. Hyg., 43: 116-123, 1990.
10. FERREIRA, A.W. - Inmunodiagnostico de la Malaria. In: ANTUŃANO, F.J. & SCHMUŃIS, G., ed. Diagnostico de la Malaria. Washington, Organización Panamericana de la Salud, 1988. p. 65-75. (Publicación Cientifica nş 512).
11. FERREIRA, A.W. & SANCHEZ, M.C.A. - Human malaria: standardization and optimization of serological tests for individual diagnosis and epidemiological surveys. Rev. Inst. Med. trop. S. Paulo, 30: 137-146, 1988.
12. FORTIER, B.; DELPLACE, P.; DUBREMETZ, J.F.; AJANA, F. & VERNES, A. - Enzyme immunoassay for detection of antigen in acute Plasmodium falciparum malaria. Europ. J. clin. Microbiol, 6: 569-598, 1987.
13. HOLMES, M.; BJORKMAN, A. & FRANZEN, L. - Diagnosis of Plasmodium falciparum infection by spot hybridization assay; specificity, sensitivity and field applicability. Bull. Wld. Hlth. Org., 64: 579-585, 1986.
14. KHUSMITH, S.; THARAVANIJ, S.; CHONGSA-NGUAN, M.; VEJVONGVARN, C. & KASEMSUTH, R. - Field applications of an immunoradiometric assay for the detection of Plasmodium falciparum antigen in a populations in a malaria-endemic area in Thailand. Amer. J. trop. Med. Hyg., 38: 3-6, 1988.
15. KORPELA, K.; LAAKSONEN, M.; KALLIO, A. et al. - Detection of Plasmodium falciparum DNA in a microtitration plate-based hybridization test. FEMS Microbiol. Lett., 90: 173-178, 1992.
16. LAL, A.A.; CHANGKASIRI, S.; HOLLINGDALE, M.R. & McCUTCHAN, T.F. - Ribosomal RNA - based diagnosis of Plasmodium falciparum malaria. Molec. Biochem. Parasit., 36: 67-72, 1989.
17. LONG, G.W.; JONES, T.R.; RICKMAN, L.S.; TRIMMER, R. & HOFFMAN, S.L. - Acridine-orange detection of Plasmodium falciparum malaria: relationship between sensitivity and optical configuration. Amer. J. trop. Med. Hyg., 44: 402-405, 1991.
18. LOWRY, O.H.; ROSEBROUGH, D.; FARR, C. & RANDAL, R.J. - Protein measure with the phenol reagent. J. Biochem., 193: 265-275, 1981.
19. MACKEY, L.; McGREGOR, I.A. & LAMBERT, P.H. - Diagnosis of Plasmodium falciparum infection using a solid-phase radioimmu-noassay for the detection of malaria antigens. Bull. Wld. Hlth. Org., 58: 439-444, 1980.
20. MACKEY, L.J.; McGREGOR, I.A.; PAOUNOVA, N. & LAMBERT, P.H. - Diagnosis of Plasmodium falciparum infection in man: detection of parasite antigens by ELISA. Bull. Wld. Hlth. Org., 60: 69-75, 1982.
21. PARZY, D.; RAPHENON, B.; MARTET, G. et al. - Quantitative Buffy Coat Test (QBC Test) Monofluo Kit Falciparum. Comparative values in the rapid diagnosis of malaria. Med. trop. (Mars.), 50: 97-101, 1990.
22. PROU, O. - Mise en évidence de Plasmodium falciparum par immunofluorescence indirectale a l'aide d'anticorps monoclonaux murins spécifiques. Rev. Transf. Immunohématol., 28: 659-670, 1985.
23. RICKMAN, L.S.; OBERST, R.; SANGALANG, R. et al. - Rapid diagnosis of malaria by acridine orange staining of centrifuged parasites. Lancet, 1: 68-71, 1989.
24. SPIELMAN, A.; PERRONE, J.B.; TEKLEHAIMANOT, A. et al. - Malaria diagnosis by direct observation of centrifuged samples of blood. Amer. J. trop. Med. Hyg., 39: 337-342, 1988.
25. THARAVANIJ, S. - New developments in malaria diagnostic techniques. Review. Southeast Asian J. trop. Med. publ. Hlth., 21: 3-21, 1990.
26. TRAGER, W. & JENSEN, J.B. - Human malaria parasites in continuous culture. Science, 193: 673-675, 1976.
27. WILSON, M.B. & NAKANE, P.K. - Recent developments in the periodate method of conjugating horseradish peroxidase (HRPO) to antibodies. In: KNAPP, W.; HOLUBAR, K. & WICK, G., ed. Immunofluorescence and related staining techniques. Amsterdam, Elsevier, North Holland Biomedical Press, 1978. p. 215-221.
28. WORLD HEALTH ORGANIZATION - Ministerial Conference on Malaria. Amsterdan. Weekly epidem. Rec., 67: 349-356, 1992.

Address for correspondence:

Sandra do Lago Moraes de Ávila

Instituto de Medicina Tropical de São Paulo. Faculdade de Medicina da Universidade de São Paulo

Av. Dr. Enéas de Carvalho Aguiar, 470

05403-000 São Paulo, SP, Brasil

(1

) The data presented in this work are part of a Ph thesis.

Publication Dates

Publication in this collection
20 Sept 2006
Date of issue
Apr 1994

History

Accepted
19 Jan 1994
Received
29 July 1993

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

[1] 1. AVILA, S.L.M.; BARNWELL, J.; NUSSENZWEIG, R.S.; ALECRIM, M.G. & FERREIRA, A.W. - Evaluation of the results of the IFI test for Plasmodia detection in patients with malaria. Possibility of application in the blood donors screening. Rev. Soc. bras. Med. trop., 25, (suppl. 2): 64, 1992.

[2] 2. AVILA, S.L.M.; MACHADO, M.; LEANDRO, M.C. et al. - Definition of serological profile in the human malaria in the sera of patients with acute and chronic disease by Western Blotting. Rev. Soc. bras. Med. trop., 25, (suppl.. 1): 6, 1992.

[3] 3. BARKER JR, R.H.; SUE BSAENG, L.; ROONEY, W. & WIRTH, D.F. - Detection of Plasmodium falciparum infection in human patients: a comparison of the DNA probe method to microscopic diagnosis. Amer. J. trop. Med. Hyg., 41: 266-272, 1989.

[4] 4. BASSET, M.T.; TAYLOR, P.; BVIRAKARE, J.; CHITEKA, F. & GOVERE, E. - Clinical diagnosis of malaria: can we improve? J. trop. Med. Hyg., 94: 65-69, 1991.

[5] 5. BRUCE-CHWATT, L.J. - From Laveran's discovery to DNA probes: new trends in diagnosis of malaria. Lancet, 2: 1509-1511, 1987.

[6] 6. BRUCE-CHWATT, L.J. - Imported malaria: an univited guest. Brit. med. Bull., 38: 179-185, 1982.

[7] 7. BRUCE-CHWATT, L.J. - Transfusional malaria revisited. Trop. Dis. Bull., 79: 827-840, 1982.

[8] 8. CARVALHO, M.C.R.; SANCHEZ, M.C.A.; QUARTIER, V.P. et al. - Transfusional malaria. Risk of contamination in non endemic areas. Rev. Inst. Med. trop. S. Paulo, 31 (supl. 7): S23, 1989.

[9] 9. DUBARRY, M.; LUILIER, M.; MALOT, N. et al. - Enzyme immunoassays for detection of malarial antigens in human plasmas by Plasmodium falciparum monoclonal antibodies. Amer. J. trop. Med. Hyg., 43: 116-123, 1990.

[10] 10. FERREIRA, A.W. - Inmunodiagnostico de la Malaria. In: ANTUŃANO, F.J. & SCHMUŃIS, G., ed. Diagnostico de la Malaria. Washington, Organización Panamericana de la Salud, 1988. p. 65-75. (Publicación Cientifica nş 512).

[11] 11. FERREIRA, A.W. & SANCHEZ, M.C.A. - Human malaria: standardization and optimization of serological tests for individual diagnosis and epidemiological surveys. Rev. Inst. Med. trop. S. Paulo, 30: 137-146, 1988.

[12] 12. FORTIER, B.; DELPLACE, P.; DUBREMETZ, J.F.; AJANA, F. & VERNES, A. - Enzyme immunoassay for detection of antigen in acute Plasmodium falciparum malaria. Europ. J. clin. Microbiol, 6: 569-598, 1987.

[13] 13. HOLMES, M.; BJORKMAN, A. & FRANZEN, L. - Diagnosis of Plasmodium falciparum infection by spot hybridization assay; specificity, sensitivity and field applicability. Bull. Wld. Hlth. Org., 64: 579-585, 1986.

[14] 14. KHUSMITH, S.; THARAVANIJ, S.; CHONGSA-NGUAN, M.; VEJVONGVARN, C. & KASEMSUTH, R. - Field applications of an immunoradiometric assay for the detection of Plasmodium falciparum antigen in a populations in a malaria-endemic area in Thailand. Amer. J. trop. Med. Hyg., 38: 3-6, 1988.

[15] 15. KORPELA, K.; LAAKSONEN, M.; KALLIO, A. et al. - Detection of Plasmodium falciparum DNA in a microtitration plate-based hybridization test. FEMS Microbiol. Lett., 90: 173-178, 1992.

[16] 16. LAL, A.A.; CHANGKASIRI, S.; HOLLINGDALE, M.R. & McCUTCHAN, T.F. - Ribosomal RNA - based diagnosis of Plasmodium falciparum malaria. Molec. Biochem. Parasit., 36: 67-72, 1989.

[17] 17. LONG, G.W.; JONES, T.R.; RICKMAN, L.S.; TRIMMER, R. & HOFFMAN, S.L. - Acridine-orange detection of Plasmodium falciparum malaria: relationship between sensitivity and optical configuration. Amer. J. trop. Med. Hyg., 44: 402-405, 1991.

[18] 18. LOWRY, O.H.; ROSEBROUGH, D.; FARR, C. & RANDAL, R.J. - Protein measure with the phenol reagent. J. Biochem., 193: 265-275, 1981.

[19] 19. MACKEY, L.; McGREGOR, I.A. & LAMBERT, P.H. - Diagnosis of Plasmodium falciparum infection using a solid-phase radioimmu-noassay for the detection of malaria antigens. Bull. Wld. Hlth. Org., 58: 439-444, 1980.

[20] 20. MACKEY, L.J.; McGREGOR, I.A.; PAOUNOVA, N. & LAMBERT, P.H. - Diagnosis of Plasmodium falciparum infection in man: detection of parasite antigens by ELISA. Bull. Wld. Hlth. Org., 60: 69-75, 1982.

[21] 21. PARZY, D.; RAPHENON, B.; MARTET, G. et al. - Quantitative Buffy Coat Test (QBC Test) Monofluo Kit Falciparum. Comparative values in the rapid diagnosis of malaria. Med. trop. (Mars.), 50: 97-101, 1990.

[22] 22. PROU, O. - Mise en évidence de Plasmodium falciparum par immunofluorescence indirectale a l'aide d'anticorps monoclonaux murins spécifiques. Rev. Transf. Immunohématol., 28: 659-670, 1985.

[23] 23. RICKMAN, L.S.; OBERST, R.; SANGALANG, R. et al. - Rapid diagnosis of malaria by acridine orange staining of centrifuged parasites. Lancet, 1: 68-71, 1989.

[24] 24. SPIELMAN, A.; PERRONE, J.B.; TEKLEHAIMANOT, A. et al. - Malaria diagnosis by direct observation of centrifuged samples of blood. Amer. J. trop. Med. Hyg., 39: 337-342, 1988.

[25] 25. THARAVANIJ, S. - New developments in malaria diagnostic techniques. Review. Southeast Asian J. trop. Med. publ. Hlth., 21: 3-21, 1990.

[26] 26. TRAGER, W. & JENSEN, J.B. - Human malaria parasites in continuous culture. Science, 193: 673-675, 1976.

[27] 27. WILSON, M.B. & NAKANE, P.K. - Recent developments in the periodate method of conjugating horseradish peroxidase (HRPO) to antibodies. In: KNAPP, W.; HOLUBAR, K. & WICK, G., ed. Immunofluorescence and related staining techniques. Amsterdam, Elsevier, North Holland Biomedical Press, 1978. p. 215-221.

[28] 28. WORLD HEALTH ORGANIZATION - Ministerial Conference on Malaria. Amsterdan. Weekly epidem. Rec., 67: 349-356, 1992.