IgY antibody and human neurocysticercosis: a novel approach on immunodiagnosis using Taenia crassiceps hydrophobic antigens

Parasitology ◽  
2019 ◽  
Vol 147 (2) ◽  
pp. 240-247 ◽  
Author(s):  
Guilherme M. P. Carrara ◽  
Gabriela B. Silva ◽  
Lucas S. Faria ◽  
Daniela S. Nunes ◽  
Vanessa S. Ribeiro ◽  
...  
Keyword(s):  
Immune Complexes ◽  
Sandwich Elisa ◽  
Taenia Solium ◽  
Antibody Test ◽  
Egg Yolk ◽  
Enzyme Linked Immunosorbent Assay ◽  
Taenia Crassiceps ◽  
Novel Approach ◽  
Human Sera ◽  
Diagnostic Values

AbstractHuman neurocysticercosis (NCC) is a worldwide neglected disease caused by Taenia solium metacestode and responsible for various complications and neurological disorders. This study aimed to evaluate the use of specific immunoglobulin Y (IgY) produced by laying hens immunized with a hydrophobic fraction of Taenia crassiceps metacestodes (hFTc) in NCC diagnosis. Egg yolk IgY antibodies were fractionated, purified and characterized. Enzyme-linked immunosorbent assay (ELISA) was carried out to evaluate the production kinetics and avidity maturation of anti-hFTc IgY antibodies throughout the IgY obtention process. Antigen recognition tests were carried out by Western blotting and immunofluorescence antibody test using purified and specific anti-hFTc IgY antibodies for detection of parasitic antigens of T. crassiceps and T. solium metacestodes. Sandwich ELISA was performed to detect circulating immune complexes formed by IgG and parasitic antigens in human sera. The results showed high diagnostic values (93.2% sensitivity and 94.3% specificity) for immune complexes detection in human sera with confirmed NCC. In conclusion, specific IgY antibodies produced from immunized hens with hFTc antigens were efficient to detect T. solium immune complexes in human sera, being an innovative and potential tool for NCC immunodiagnosis.

Use of polyclonal IgY antibodies to detect serum immune complexes in patients with active hookworm infection

Parasitology ◽  
2020 ◽  
Vol 147 (6) ◽  
pp. 715-720
Author(s):  
Dayane Costa de Souza ◽  
Lucas Silva de Faria ◽  
José Eduardo Neto de Sousa ◽  
Camila de Almeida Lopes ◽  
Vanessa da Silva Ribeiro ◽  
...  
Keyword(s):  
Immune Complexes ◽  
Protein Complexes ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sandwich Elisa ◽  
Sodium Dodecyl ◽  
Egg Yolk ◽  
Diagnostic Value ◽  
Enzyme Linked Immunosorbent Assay ◽  
Hookworm Infection ◽  
Serum Samples

AbstractDefinitive diagnosis of hookworm infection is usually based on the microscopic detection of eggs in a stool sample; however, several cases display a low or irregular egg output. Serodiagnosis can be a useful tool to identify these cases, but conventional tests do not differentiate past from active infections. The aim of this study was to obtain and apply egg yolk polyclonal immunoglobulin (IgY) antibodies to detect immune complexes (ICs) in serum samples from patients infected with hookworm. Hens were immunized with Ancylostoma ceylanicum saline extract, their eggs were collected and then IgY antibodies were extracted and purified. Antibody purity was tested by 12% sodium dodecyl sulphate polyacrylamide gel electrophoresis and specificity was assessed by immunoblotting and immunofluorescence. IgY production was evaluated by kinetics enzyme-linked immunosorbent assay (ELISA). Sandwich ELISA tested the ability of IgY to detect ICs in serum samples, from which diagnostic parameters were calculated. Antibody responses increased steadily from day 7 to 42. In the immunoblotting assay, IgY recognized two protein complexes. The immunofluorescence assay showed no staining in control samples. The sandwich ELISA presented a very high diagnostic value, with a sensitivity of 90% and a specificity of 86.7%. Our pioneer strategy highlights the potential use of egg yolk IgY as a diagnostic test to detect active hookworm infection.

scholarly journals Frequency of serum anti-cysticercus antibodies in the population of a rural Brazilian community (Cássia dos Coqueiros, SP) determined by ELISA and immunoblotting using Taenia crassiceps antigens

2002 ◽  
Vol 44 (1) ◽  
pp. 7-12 ◽  
Author(s):  
Lúcia M. BRAGAZZA ◽  
Adelaide J. VAZ ◽  
Afonso D.C. PASSOS ◽  
Osvaldo M. TAKAYANAGUI ◽  
Paulo M. NAKAMURA ◽  
...  
Keyword(s):  
Public Health ◽  
Immunosorbent Assay ◽  
Taenia Solium ◽  
High Specificity ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Taenia Crassiceps ◽  
Municipal District ◽  
The Impact

Considering the impact of cysticercosis on public health, especially the neurologic form of the disease, neurocysticercosis (NC), we studied the frequency of positivity of anti-Taenia solium cysticercus antibodies in serum samples from 1,863 inhabitants of Cássia dos Coqueiros, SP, a municipal district located 80 km from Ribeirão Preto, an area considered endemic for cysticercosis. The 1,863 samples were tested by enzyme linked immunosorbent assay (ELISA) using an antigenic extract from Taenia crassiceps vesicular fluid (Tcra). The reactive and inconclusive ELISA samples were tested by immunoblotting. Of the 459 samples submitted to immunoblotting, 40 were strongly immunoreactive to the immunodominant 18 and 14 kD peptides. Considering the use of immunoblotting as confirmatory due to its high specificity, the anti-cysticercus serum prevalence in this population was 2.1%.

Anti-Ascaris suum immunoglobulin Y as a novel biotechnological tool for the diagnosis of human ascariasis

2019 ◽  
Vol 94 ◽  
Author(s):  
C.A. Lopes ◽  
L.S. de Faria ◽  
J.E.N. de Sousa ◽  
I.P. Borges ◽  
R.P. Ribeiro ◽  
...  
Keyword(s):  
Immune Complexes ◽  
Ascaris Suum ◽  
Sandwich Elisa ◽  
Adult Life ◽  
Disease Diagnosis ◽  
Diagnostic Methods ◽  
Enzyme Linked Immunosorbent Assay ◽  
High Avidity ◽  
Immunoglobulin Y ◽  
Saline Extract

Abstract Human ascariasis is a neglected tropical disease of great relevance to public health and is considered the most frequent helminthiasis in poor regions. Accurately diagnosing this parasite has been challenging due to limitations of current diagnostic methods. Immunoglobulin Y (IgY) technology is a very effective alternative for the production of highly specific and profitable antibodies. This study aimed to produce and apply anti-Ascaris suum IgY antibodies in the immunodiagnosis of human ascariasis. Five immunizations comprising total saline extract from A. suum adult life forms were given at 14-day intervals to Gallus gallus domesticus hens of the Isa Brown line. Eggs and blood samples were collected weekly and fortnightly, respectively, to monitor the production of antibodies. The specificity of antibodies was confirmed by dot-blot, kinetic enzyme-linked immunosorbent assay (ELISA), avidity ELISA, immunoblotting and indirect immunofluorescence antibody tests. The application for disease diagnosis was performed through the detection of immune complexes in human serum samples by sandwich ELISA. Peaks of IgY anti-A. suum production occurred at weeks 6 and 8. IgY showed high avidity levels after the second dose of immunization, ranging from 64% to 93%, with a mean avidity index of 78.30%. Purified IgY recognized 12 bands of proteins from A. suum saline extract. Eggs, the uterine portion and cuticles of A. suum female adult are reactive in immunofluorescence. The detection of immune complexes showed diagnostic values of 80% sensitivity and 90% specificity. In conclusion, specific IgY have been shown to be a potential immunodiagnostic tool with promising future applications in human ascariasis.

scholarly journals Leishmania major-Like Antigen for Specific and Sensitive Serodiagnosis of Human and Canine Visceral Leishmaniasis

2002 ◽  
Vol 9 (6) ◽  
pp. 1361-1366 ◽  
Author(s):  
Rosângela Barbosa-de-Deus ◽  
Marcos Luíz dos Mares-Guia ◽  
Adriane Zacarias Nunes ◽  
Kátia Morais Costa ◽  
Roberto Gonçalves Junqueira ◽  
...  
Keyword(s):  
Visceral Leishmaniasis ◽  
Rheumatoid Factor ◽  
Chagas Disease ◽  
Leishmania Major ◽  
Antibody Test ◽  
Cross Reactivity ◽  
Enzyme Linked Immunosorbent Assay ◽  
Parasitological Diagnosis ◽  
Human Sera ◽  
Indirect Immunofluorescent

ABSTRACT An antigen (LMS) prepared from Leishmania major-like promastigotes was used in an enzyme-linked immunosorbent assay (ELISA) for the diagnosis of human and dog visceral leishmaniasis. The results were compared with those from the indirect immunofluorescent antibody test (IFAT). A total of 1,822 canine sera were tested, including sera from dogs with visceral leishmaniasis, transmissible venereal tumors, ehrlichiosis, rickettsiosis, or Chagas' disease and sera from healthy dogs. The antigen was also tested with 227 samples of human sera, including sera from patients with visceral, cutaneous, or diffuse cutaneous leishmaniasis and from noninfected individuals, as well as sera from patients with Chagas' disease, toxoplasmosis, rickettsiosis, hepatitis B, schistosomiasis, ascaridiasis, malaria, rheumatoid factor, leprosy and rheumatoid factor, tuberculosis, or leprosy. All dogs and all human patients had a clinical and/or serological and/or parasitological diagnosis. For detecting antibodies in sera from dogs with leishmaniasis, the antigen showed a sensitivity of 98%, specificity of 95%, and concordance of 93% and when used for detecting antibodies in human sera presented a sensitivity of 92%, specificity of 100%, and concordance of 92%. Comparison between ELISA and IFAT demonstrated that ELISA using the LMS antigen yielded more reliable results than IFAT. The LMS antigen displayed no cross-reactivity with sera from patients or dogs that had any of the other diseases tested.

scholarly journals Specific Taenia crassiceps and Taenia solium Antigenic Peptides for Neurocysticercosis Immunodiagnosis Using Serum Samples

2000 ◽  
Vol 38 (1) ◽  
pp. 146-151
Author(s):  
Ednéia Casagranda Bueno ◽  
Adelaide José Vaz ◽  
Luís Dos Ramos Machado ◽  
JoséAntônio Livramento ◽  
Sílvia Regina Mielle
Keyword(s):  
Taenia Solium ◽  
Severe Infection ◽  
Enzyme Linked Immunosorbent Assay ◽  
Control Group ◽  
Antigenic Peptides ◽  
Serum Samples ◽  
Immunoblotting Analysis ◽  
Taenia Crassiceps ◽  
The Central Nervous System ◽  
Paired Serum

ABSTRACT Neurocysticercosis (NC), i.e., the presence of the larval form of Taenia solium in tissues, is the most frequent and severe infection involving the central nervous system. Paired serum and cerebrospinal fluid (CSF) samples from patients with NC, CSF and serum samples from a control group, and serum samples from patients with other parasitoses were studied by enzyme-linked immunosorbent assay (ELISA) and by immunoblotting with Taenia crassiceps vesicular fluid antigen (Tcra) and Taenia solium total saline antigen (Tso) for the detection of immunoglobulin G antibodies. ELISAs carried out with the Tso and Tcra antigens showed 94.1 and 95.6% sensitivities, respectively, for the detection of antibodies in CSF and 70.6% and 91.2% sensitivities, respectively, for the detection of antibodies in serum, with 100% specificity for the detection of antibodies in CSF and 80% specificity for the detection of antibodies in serum for both antigens. On the basis of the reactivities of the peptides in the samples analyzed, the peptides of ≤23, 39, 85 to 77, and 97 kDa were found to be Tso specific by immunoblotting and the peptides of ≤62, 74, 109, 121, and 131 kDa were found to be Tcra specific. Tests with Tcra extract had higher sensitivities and more homogeneous results and permitted us to obtain the parasites easily. We suggest the use of Tcra ELISA for the study of serum and confirmation of the results for sera positive by an immunoblotting analysis in which specific peptides (e.g., peptides of 19 to 13 kDa) are detected.

scholarly journals Anti-Taenia solium metacestodes antibodies in serum from blood donors from four cities of Triângulo Mineiro area, Minas Gerais, Brazil, 1995

2002 ◽  
Vol 44 (4) ◽  
pp. 229-231 ◽  
Author(s):  
Elisângela de Paula SILVEIRA-LACERDA ◽  
Eleuza Rodrigues MACHADO ◽  
Sílvio César de Freitas ARANTES ◽  
Julia Maria COSTA-CRUZ
Keyword(s):  
Blood Donors ◽  
Taenia Solium ◽  
Antibody Test ◽  
Minas Gerais ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serological Survey ◽  
Igg Antibodies ◽  
Specific Igg ◽  
Indirect Fluorescence Antibody ◽  
Specific Igg Antibodies

Serological survey was performed to detect IgG antibodies anti-Taenia solium metacestodes in blood donors of Hemocentro Regional de Uberlândia, Minas Gerais, Brazil. A total of 1133 sera from blood donors coming from four cities of Triângulo Mineiro area were analyzed by the indirect fluorescence antibody test (IFAT) and the enzyme linked immunosorbent assay (ELISA). Specific IgG antibodies were found in 5.6% of the studied population, showing differences in the positive rates according to their origin: Araguari (13.5%), Tupaciguara (5.0%), Monte Alegre de Minas (4.8%) and Uberlândia (4.7%). The results indicate the probable endemicity of cysticercosis in this population.

Evaluation of a monoclonal antibody-based competition ELISA for the diagnosis of human hydatidosis

Parasitology ◽  
1992 ◽  
Vol 104 (2) ◽  
pp. 357-361 ◽  
Author(s):  
D. Liu ◽  
M. W. Lightowlers ◽  
M. D. Rickard
Keyword(s):  
Taenia Solium ◽  
Disease Diagnosis ◽  
Enzyme Linked Immunosorbent Assay ◽  
Competition Elisa ◽  
Combined Use ◽  
Highly Sensitive ◽  
Human Sera ◽  
Normal Human ◽  
Conventional Antibody ◽  
Antibody Competition

SUMMARYAn antibody competition enzyme-linked immunosorbent assay (ELISA) using 4 different monoclonal antibodies (MAb) raised against major antigens (Antigen 5 and Antigen B) of Echinococcus granulosus was evaluated for the diagnosis of human hydatidosis. The competition assay, using anti-Ag5 MAb 24.14, detected specific antibodies in 70% (131/188) of sera from patients with surgically confirmed E. granulosus infection and 38·5% (10/26) of sera from patients with E. multilocularis infection. None of the sera from patients with Taenia solium cysticercosis (10), T. saginata (2), filariasis (22), strongyloidiasis (19), fascioliasis (4), bilharziasis (4) and amoebiasis (2) tested were positive using a cut-off point established through reaction between MAb 24.14 and normal human sera. The combined use of the MAb 24.14-based competition ELISA with the conventional antibody-binding assay provides a highly sensitive (92·8%) and specific screening system for human hydatid disease diagnosis.

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