Epitope mapping of recombinant Leishmania donovani virulence factor A2 (recLdVFA2) and canine leishmaniasis diagnosis using a derived synthetic bi-epitope
| Autor: | Fernanda Costal-Oliveira, Ricardo Toshio Fujiwara, Carlos Chávez-Olórtegui, Oscar Bruna-Romero, Eric Henrique Roma, Cristina Monerat Toledo-Machado, T.M. Mendes, Daniella Castanheiras Bartholomeu, Lucas de Carvalho Dhom-Lemos |
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| Jazyk: | angličtina |
| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Proteomics B Cells Physiology Protozoan Proteins Antibodies Protozoan Biochemistry Epitope White Blood Cells 0302 clinical medicine Animal Cells Immune Physiology Zoonoses Canine leishmaniasis Medicine and Health Sciences Dog Diseases Enzyme-Linked Immunoassays Leishmania infantum Leishmaniasis Mammals Immune System Proteins biology lcsh:Public aspects of medicine Chemical Synthesis Infectious Diseases Vertebrates Epitopes B-Lymphocyte Leishmaniasis Visceral Cellular Types Research Article Neglected Tropical Diseases lcsh:Arctic medicine. Tropical medicine Biosynthetic Techniques Virulence Factors lcsh:RC955-962 Immune Cells 030231 tropical medicine Immunology Leishmania donovani Antigens Protozoan Enzyme-Linked Immunosorbent Assay Research and Analysis Methods Peptide Mapping Antibodies 03 medical and health sciences Dogs Antigen medicine Parasitic Diseases Animals Serologic Tests Amino Acid Sequence Immunoassays Molecular Biology Techniques Antibody-Producing Cells Peptide Synthesis Molecular Biology Protozoan Infections Blood Cells Gene Mapping Public Health Environmental and Occupational Health Organisms Biology and Life Sciences Proteins lcsh:RA1-1270 Cell Biology medicine.disease biology.organism_classification Tropical Diseases Virology 030104 developmental biology Epitope mapping Visceral leishmaniasis Amniotes Immunologic Techniques Epitope Mapping |
| Zdroj: | PLoS Neglected Tropical Diseases, Vol 11, Iss 5, p e0005562 (2017) PLoS Neglected Tropical Diseases |
| ISSN: | 1935-2735 1935-2727 |
| Popis: | Background Leishmaniasis is one of the most important zoonotic diseases spread in Latin America. Since many species are involved in dog infection with different clinical manifestations, the development of specific diagnostic tests is mandatory for more accurate disease control and vaccine strategies. Methodology/Principal findings Seventy-five 15-mer peptides covering the sequence of recombinant Leishmania donovani virulence factor A2 (recLdVFA2) protein were prepared by Spot synthesis. Membrane-bound peptides immunoreactivity with sera from dogs immunized with recLdVFA2 and with a specific anti-recLdVFA2 monoclonal antibody allowed mapping of continuous B-cell epitopes. Five epitopes corresponding to the N-terminal region of recLdVFA2 (MKIRSVRPLVVLLVC, RSVRPLVVLLVCVAA, RPLVVLLVCVAAVLA, VVLLVCVAAVLALSA and LVCVAAVLALSASAE, region 1–28) and one located within the repetitive units (PLSVGPQAVGLSVG, regions 67–81 and 122–135) were identified. A 34-mer recLdVFA2-derived bi-epitope containing the sequence MKIRSVRPLVVLLVC linked to PLSVGPQAVGLSVG by a Gly-Gly spacer was chemically synthesized in its soluble form. The synthetic bi-epitope was used as antigen to coat ELISA plates and assayed with dog sera for in vitro diagnosis of canine visceral leishmaniasis (CVL). The assay proved to be highly sensitive (98%) and specific (99%). Conclusions/Significance Our work suggests that synthetic peptide-based ELISA strategy may be useful for the development of a sensitive and highly specific serodiagnosis for CVL or other parasitic diseases. Author summary Leishmaniasis is a neglected tropical disease being among the six endemic prioritized diseases in the world. Visceral leishmaniasis (VL) is caused by Leishmania infantum and represents a serious public health problem in Brazil. Dogs are the main source of infection in the urban area and, in Brazil, the main strategies of the Visceral Leishmaniasis Control Program are directed to control the canine reservoir (serological survey and euthanasia of dogs which present reactive serum). In general, diagnosis of canine visceral leishmaniasis (CVL) has been presented as a problem for Brazilian public health services. The issue should be attributed mainly to the following factors: 1- range of similar clinical signs observed in other infectious diseases that affect dogs; 2- large percentage of asymptomatic or oligosymptomatic dogs; 3- nonspecific histopathological changes; 4- nonexistence of a diagnostic test 100% specific and sensitive. In this work, we developed a synthetic bi-epitope peptide as an antigen for immunodiagnostic ELISA to detect CVL. The biepitope used for ELISA assay accurately distinguish (98% sensitivity and 99% specificity) CVL dogs sera from non-infected dogs sera. |
| Databáze: | OpenAIRE |
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