Polyclonal antibodies for the detection of Trypanosoma cruzi circulating antigens
| Autor: | Sofía Astupiña-Figueroa, Noelia Angulo, Manuela Verastegui, Remo A. Gonza, Edith S. Málaga-Machaca, Ada del Carpio-Sanz, Alessandra Romero-Ramirez, Fernando Recuenco, Maritza Calderon, Holger Mayta, Rosina Camargo, Liliam A. Barrueta-Soria, Robert H. Gilman, Cinthya J. Lovon-Luque, Ines Cabello, Alejandro Florentini |
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| Jazyk: | angličtina |
| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Life Cycles Physiology Antibodies Protozoan Urine Protozoology Pathology and Laboratory Medicine Biochemistry law.invention Serology 0302 clinical medicine law Immune Physiology Medicine and Health Sciences Enzyme-Linked Immunoassays Protozoans Immune System Proteins biology medicine.diagnostic_test lcsh:Public aspects of medicine Eukaryota 3. Good health Body Fluids Infectious Diseases Recombinant DNA Protozoan Life Cycles Trypanosoma cruzi/immunology Rabbits Antibody Anatomy Camelids New World purl.org/pe-repo/ocde/ford#3.03.06 [https] Research Article Neglected Tropical Diseases Chagas disease Trypanosoma lcsh:Arctic medicine. Tropical medicine lcsh:RC955-962 Trypanosoma cruzi 030231 tropical medicine Immunology Antigens Protozoan Enzyme-Linked Immunosorbent Assay Research and Analysis Methods Microbiology Antibodies 03 medical and health sciences Antigen Western blot medicine Parasitic Diseases Animals Chagas Disease Serologic Tests Immunoassays Protozoan Infections Chagas Disease/diagnosis Serologic Tests/methods Enzyme-Linked Immunosorbent Assay/methods Antibodies Protozoan/immunology Public Health Environmental and Occupational Health Organisms Biology and Life Sciences Proteins lcsh:RA1-1270 Trypomastigotes biology.organism_classification medicine.disease Tropical Diseases Virology Parasitic Protozoans 030104 developmental biology Antigens Protozoan/blood/urine Polyclonal antibodies biology.protein Immunologic Techniques Chickens Developmental Biology |
| Zdroj: | PLOS NEGLECTED TROPICAL DISEASES PLoS Neglected Tropical Diseases PLoS Neglected Tropical Diseases, Vol 11, Iss 11, p e0006069 (2017) |
| Popis: | Background Detection of Trypanosoma cruzi antigens in clinical samples is considered an important diagnostic tool for Chagas disease. The production and use of polyclonal antibodies may contribute to an increase in the sensitivity of immunodiagnosis of Chagas disease. Methodology/Principal findings Polyclonal antibodies were raised in alpacas, rabbits, and hens immunized with trypomastigote excreted-secreted antigen, membrane proteins, trypomastigote lysate antigen and recombinant 1F8 to produce polyclonal antibodies. Western blot analysis was performed to determine specificity of the developed antibodies. An antigen capture ELISA of circulating antigens in serum, plasma and urine samples was developed using IgY polyclonal antibodies against T. cruzi membrane antigens (capture antibody) and IgG from alpaca raised against TESA. A total of 33 serum, 23 plasma and 9 urine samples were analyzed using the developed test. Among serum samples, compared to serology, the antigen capture ELISA tested positive in 55% of samples. All plasma samples from serology positive subjects were positive in the antigen capture ELISA. All urine positive samples had corresponding plasma samples that were also positive when tested by the antigen capture ELISA. Conclusions Polyclonal antibodies are useful for detection of circulating antigens in both the plasma and urine of infected individuals. Detection of antigens is direct evidence of the presence of the parasite, and could be a better surrogate of current infection status. Author summary Current diagnosis of Chagas disease is still cumbersome. Diagnosis is based on antibody detection with at least two assays of distinct mechanisms. If a discrepancy exists, a third assay must be performed. However, detection of antibodies is not indicative of current infection. Molecular-based techniques such as qPCR have been used for diagnosis and as a gold standard in the demonstration of therapeutic failure, but availability of genomic material depends on the presence of parasites in the bloodstream. Detection of parasite-derived antigens represents a better alternative for diagnosis, as several proteins are secreted/excreted by the parasites and may be detected in blood and in the urine of infected individuals. This study describes the development of polyclonal antibodies raised against different Trypanosoma cruzi antigens and their applicability for the diagnosis of Chagas disease using the widely-used ELISA format. |
| Databáze: | OpenAIRE |
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