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Usefulness of gel filtration fraction as potential biomarker for neurocysticercosis in serum: towards a new diagnostic tool

  • D. S. NUNES (a1), H. T. GONZAGA (a1), V. S. RIBEIRO (a1) (a2), J. P. CUNHA-JÚNIOR (a1) and J. M. COSTA-CRUZ (a1)...

Summary

There is an increasing interest in improving neurocysticercosis (NCC) diagnosis through the search of new and alternative antigenic sources, as those obtained from heterologous antigens. The aim of this study was to obtain potential biomarkers for NCC diagnosis after gel filtration chromatography [gel filtration fraction (GFF)] from the total saline extract (SE) from Taenia saginata metacestodes, followed by protein identification and application in immunodiagnostic. SE and GFF proteic profiles were characterized in gel electrophoresis, and diagnostic performance was verified by testing 160 serum samples through enzyme-linked immunosorbent assay and immunoblotting. Sensitivity (Se), specificity (Sp) and other diagnostic parameters were calculated. Polypeptides of interest in the diagnosis of human NCC present at GFF were analysed by mass spectrometry (MS) and B-cell epitopes were predicted. GFF had the best diagnostic parameters: Se 93·3%; Sp 93%; AUC 0·990; LR+ = 13·42 and LR− = 0·07, and proved to be useful reacting with serum samples in immunoblotting. Proteic profile ranged from 64 to 68 kDa and enolase and calcium binding protein calreticulin precursor were identified after MS. The enolase and calcium-binding protein calreticulin precursor showed 18 and 10 predicted B-cell epitopes, respectively. In conclusion we identified important markers in the GFF with high efficiency to diagnose NCC.

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Corresponding author

*Corresponding author: Laboratório de Diagnóstico de Parasitoses, Departamento de Imunologia, Microbiologia e Parasitologia, Instituto de Ciências Biomédicas, Universidade Federal de Uberlândia, Uberlândia, MG, Brasil. Avenida Pará 1720, 38400-902, Uberlândia, MG, Brazil. E-mail: costacruz@ufu.br

References

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Arriola, C. S., Gonzalez, A. E., Gomez-Puerta, L. A., Lopez-Urbina, M. T., Garcia, H. H. and Gilman, R. H. (2014). New insights in cysticercosis transmission. PLoS Neglected Tropical Diseases 10, e3247.
Assana, E., Kanobana, K., Tume, C. B., Zoli, P. A., Nguekam, J. P., Geerts, S., Berkvens, D. and Dorny, P. (2007). Isolation of a 14 kDa antigen from Taenia solium cyst fluid by HPLC and its evaluation in enzyme linked immunosorbent assay for diagnosis of porcine cysticercosis. Research in Veterinary Science 82, 370376.
Baermann, G. (1917). Eine einfache Methode zur Auffindung von Ankylostomum (Nematoden) Larven in Erdproben Mededeel mit H Geneesk Laboratories Weltevreden, Feestbundel, Batavia, pp. 4147.
Barcelos, I. S., Mineo, J. R., Oliveira-Silva, D. A., Ferreira, M. S., Moura, L. P., Biondi, G. F. and Costa-Cruz, J. M. (2001). Detection of IgG in cerebrospinal fluid for diagnosis of neurocysticercosis: evaluation of saline and SDS extracts from Taenia solium and Taenia crassiceps metacestodes by ELISA and immunoblot assay. Tropical Medicine and International Health 6, 219226.
Barcelos, I. S., Moura, L. P., Costa, V. P., Ferreira, M. S. and Costa-Cruz, J. M. (2007). Taenia solium metacestode immunodominant peptides recognized by IgG antibodies in cerebrospinal fluid and serum paired samples from patients with active and inactive neurocysticercosis. Memórias do Instituto Oswaldo Cruz 102, 713717.
Bradford, M. A. (1976). Rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry 72, 248254.
Carpio, A. and Romo, M. L. (2015). Multifactorial basis of epilepsy in patients with neurocysticercosis. Epilepsia 56, 973974.
Cayir, E., Erdemir, A., Ozkan, E., Topuzogullari, M., Bolat, Z. B., Akat, A. and Turgut-Balik, D. (2014). Cloning of intron-removed enolase gene and expression, purification, kinetic characterization of the enzyme from Theileria annulata . Molecular Biotechnology 56, 689696.
Chen, N., Yuan, Z. G., Xu, M. J., Zhou, D. H., Zhang, X. X., Zhang, Y. Z., Wang, X. W., Yan, C., Lin, R. Q. and Zhu, X. Q. (2012). Ascaris suum enolase is a potential vaccine candidate against ascariasis. Vaccine 30, 34783482.
Coral-Almeida, M., Gabriël, S., Abatih, E. N., Praet, N., Benitez, W. and Dorny, P. (2015). Taenia solium human cysticercosis: a systematic review of sero-epidemiological data from endemic zones around the world. PLoS Neglected Tropical Diseases 9, e0003919.
Costa, J. M., Ferreira, A. W., Makino, M. M. and Camargo, M. E. (1982). Spinal fluid immunoenzymatic assay (ELISA) for neurocysticercosis. Revista do Instituto de Medicina Tropical de São Paulo 24, 337341.
Del Brutto, O. H. (2012). Diagnostic criteria for neurocysticercosis, revisited. Pathogens and Global Health 106, 299304.
Del Brutto, O. H. and García, H. H. (2015). Taenia solium cysticercosis – the lessons of history. Journal of the Neurological Sciences 359, 392395.
Del Brutto, O. H., Lama, J., Zambrano, M. and Del Brutto, V. J. (2014). Neurocysticercosis is a neglected microbleed mimic. A cautionary note for stroke neurologists. European Neurology 72, 306308.
Dong, W. H., Wang, T. Y., Wang, F. and Zhang, J. H. (2011). Simple, time-saving dye staining of proteins for sodium dodecyl sulfate-polyacrylamide gel electrophoresis using Coomassie blue. PLoS ONE 6, e22394.
Espíndola, N. M., Iha, A. H., Fernandes, I., Takayanagui, O. M., Machado, L. R., Livramento, J. A., Mendes-Maia, A. A., Peralta, J. M. and Vaz, A. J. (2005). Cysticercosis immunodiagnosis using 18- and 14-kilodalton proteins from Taenia crassiceps cysticercus antigens obtained by immunoaffinity chromatography. Journal of Clinical Microbiology 43, 31783184.
Gabriël, S., Johansen, M. V., Pozio, E., Smit, G. S., Devleesschauwer, B., Allepuz, A., Papadopoulos, E., Van Der Giessen, J. and Dorny, P. (2015). Human migration and pig/pork import in the European Union: what are the implications for Taenia solium infections? Veterinary Parasitology 213, 3845.
Gonçalves, F. A., Machado, G. A., Oliveira, H. B., Rezende, M. T., Mineo, J. R. and Costa-Cruz, J. M. (2010). Hydrophobic fraction of Taenia saginata metacestodes, rather than hydrophilic fraction, contains immunodominant markers for diagnosing human neurocysticercosis. Revista da Sociedade Brasileira de Medicina Tropical 43, 254259.
Gonzales, I., Rivera, J. T. and Garcia, H. H. (2016). Pathogenesis of Taenia solium taeniasis and cysticercosis. Parasite Immunology 38, 136146.
Granholm, V. and Käll, L. (2011). Quality assessments of peptide-spectrum matches in shotgun proteomics. Proteomics 11, 10861093.
Greiner, M., Sohr, D. and Göbel, P. (1995). A modified ROC analysis for the selection of cut-off values and the definition of intermediate results of serodiagnostic tests. Journal of Immunological Methods 185, 123132.
Hanley, J. A. and McNeil, B. J. (1982). The meaning and use of the area under a receiver operating characteristic (ROC) curve. Radiology 143, 2936.
Hanley, J. A. and McNeil, B. J. (1983). A method of comparing the areas under receiver operating characteristic curves derived from the same cases. Radiology 148, 839843.
Husain, N., Shukla, N., Kumar, R., Husain, M., Chaturvedi, A., Agarwal, G. G. and Gupta, R. K. (2008). ELISA in the evaluation of therapeutic response to albendazole in neurocysticercosis. Journal of Infection 56, 6573.
Jaeschke, R., Guyatt, G. H. and Sacket, D. L. (1994). Users’ guide to the medical literature. III. How to use an article about a diagnostic test. B. What are the results and will they help me in caring for my patients? The Evidence-Based Medicine Working Group. Journal of the American Medical Association 271, 703707.
Laemmli, U. K. (1970). Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227, 680685.
Lutz, A. (1919). O Schistosoma mansoni e a schistosomose segundo observações feitas no Brasil. Memórias do Instituto Oswaldo Cruz 11, 121125.
Machado, G. A., Santiago, F. M., Mineo, J. R. and Costa-Cruz, J. M. (2007). Assessment of antigenic fractions of varying hydrophobicity from Taenia solium metacestodes for the diagnosis of human neurocysticercosis. Tropical Medicine and International Health, 12, 13691376.
Madrazo, I. (2013). Overview of neurocysticercosis. World Neurosurgery 79, 262263.
Martinez, E. Z., Louzada-Neto, F. and Pereira, B. B. (2003). A curva ROC para testes diagnósticos. Cadernos de Saúde Coletiva 11, 731.
Martins-Melo, F. R., Ramos, A. N. Jr., Cavalcanti, M. G., Alencar, C. H. and Heukelbach, J. (2016). Neurocysticercosis-related mortality in Brazil, 2000–2011: epidemiology of a neglected neurologic cause of death. Acta Tropica 153, 128136.
Mendlovic, F., Ostoa-Saloma, P., Solís, C. F., Martínez-Ocaña, J., Flisser, A. and Laclette, J. P. (2004). Cloning, characterization, and functional expression of Taenia solium calreticulin. Journal of Parasitology 90, 891893.
Moraes, R. G. (1948). Contribuição para o estudo do Strongyloides stercoralis e da estrongiloidíase no Brasil. Revista de Saúde Pública 1, 507624.
Morakote, N., Nawacharoen, W., Sukonthasun, K., Thammasonthi, W. and Khamboonruang, C. (1992). Comparison of cysticercus extract, cyst fluid and Taenia saginata extract for use in ELISA for serodiagnosis of neurocysticercosis. Southeast Asian Ministers of Education Council Central Coordinating Board for Tropical Medicine and Public Health 23, 7781.
Ndimubanzi, P. C., Carabin, H., Budke, C. M., Nguyen, H., Qian, Y. J., Rainwater, E., Dickey, M., Reynolds, S. and Stoner, J. A. (2010). A systematic review of the frequency of neurocyticercosis with a focus on people with epilepsy. PLoS Neglected Tropical Diseases 4, e870.
Nunes, D. S., Ribeiro, V. S., Manhani, M. N. and Costa-Cruz, J. M. (2010). Jacalin-unbound fraction of Taenia saginata in immunodiagnosis of neurocysticercosis in human cerebrospinal fluid. Diagnostic Microbiology and Infectious Disease 68, 259264.
Nunes, D. S., Gonzaga, H. T., Ribeiro, V. S., Cunha-Junior, J. P. and Costa-Cruz, J. M. (2013). Taenia saginata metacestode antigenic fractions obtained by ion-exchange chromatography: potential source of immunodominant markers applicable in the immunodiagnosis of human neurocysticercosis. Diagnostic Microbiology and Infectious Disease 76, 3641.
O'keefe, K. A., Eberhard, M. L., Shafir, S. C., Wilkins, P., Ash, L. R. and Sorvillo, F. J. (2014). Cysticercosis-related hospitalizations in the United States, 1998–2011. The American Journal of Tropical Medicine and Hygiene 10, 140506.
Oliveira, H. B., Machado, G. A., Cabral, D. D. and Costa-Cruz, J. M. (2007). Application of Taenia saginata metacestodes as an alternative antigen for the serological diagnosis of human neurocysticercosis. Parasitology Research 101, 10071013.
Oliveira, H. B., Machado, G. A., Gonçalves-Pires, M. R. F., Moura, L. P. and Costa-Cruz, J. M. (2009). Saline extract of Taenia saginata metacestodes as alternative antigen for the immunodiagnosis of neurocysticercosis in human cerebrospinal fluid. Parasitology Research 105, 169174.
Oliveira, H. B., Machado, G. A., Mineo, J. R. and Costa-Cruz, J. M. (2010). Taenia saginata metacestode antigenic fractions without affinity to concanavalin A are an important source of specific antigens for the diagnosis of human neurocysticercosis. Clinical and Vaccine Immunology 17, 638644.
Ramajo-Hernández, A., Pérez-Sánchez, R., Ramajo-Martín, V. and Oleaga, A. (2007). Schistosoma bovis: plasminogen binding in adults and the identification of plasminogen-binding proteins from the worm tegument. Experimental Parasitology 115, 8391.
Ribeiro, V. S., Nunes, D. S., Gonzaga, H. T., Cunha-Júnior, J. P. and Costa-Cruz, J. M. (2014). Diethylaminoethyl (DEAE) binding fraction from Taenia solium metacestode improves the neurocysticercosis serodiagnosis. Parasitology Research 113, 25692575.
Ritchie, L. S. (1948). An ether sedimentation technique for rotine stool examination. Bulletin of the National Service of Army – Medical Department 8, 326334.
Rizvi, S. A., Saleh, A. M., Frimpong, H. A., Mohiy, H. M., Ahmed, J., Edwards, R. D. and Ahmed, S. S. (2016). Neurocysticercosis: a case report and brief review. Asian Pacific Journal of Tropical Medicine 9, 100102.
Shiguekawa, K. Y., Mineo, J. R., Moura, L. P. and Costa-Cruz, J. M. (2000). ELISA and western blotting tests in the detection of IgG antibodies to Taenia solium metacestodes in serum samples in human neurocysticercosis. Tropical Medicine and International Health 5, 443449.
Sotelo, J., Guerrero, V. and Rubio, F. (1985). Neurocysticercosis: a new classification based on active and inactive forms. A study of 753 cases. Archives of Internal Medicine 145, 442445.
Towbin, H., Staehelin, T. and Gordon, J. (1979). Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proceedings of the National Academy of Sciences of the United States of America 76, 43504354.
Vasoo, S. and Pritt, B. S. (2013). Molecular diagnostics and parasitic disease. Clinics in Laboratory Medicine 33, 461503.
Yang, J., Qiu, C., Xia, Y., Yao, L., Fu, Z., Yuan, C., Feng, X. and Lin, J. (2010). Molecular cloning and functional characterization of Schistosoma japonicum enolase which is highly expressed at the schistosomulum stage. Parasitology Research 107, 667677.
Youden, W. J. (1950). Index for rating diagnostic tests. Cancer 3, 3235.
Zhao, Y. and Lin, Y. H. (2010). Whole-cell protein identification using the concept of unique peptides. Genomics Proteomics Bioinformatics 8, 3341.
Zimic, M., Pajuelo, M., Rueda, D., López, C., Arana, Y., Castillo, Y., Calderón, M., Rodriguez, S., Sheen, P., Vinetz, J. M., Gonzales, A., García, H. H., Gilman, R. H. (2009). Utility of a protein fraction with cathepsin L-Like activity purified from cysticercus fluid of Taenia solium in the diagnosis of human cysticercosis. The American Journal of Tropical Medicine and Hygiene 80, 964970.

Keywords

Usefulness of gel filtration fraction as potential biomarker for neurocysticercosis in serum: towards a new diagnostic tool

  • D. S. NUNES (a1), H. T. GONZAGA (a1), V. S. RIBEIRO (a1) (a2), J. P. CUNHA-JÚNIOR (a1) and J. M. COSTA-CRUZ (a1)...

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