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.
Background
Neurocysticercosis (NCC) is a neglected tropical disease and its diagnosis is still a challenge due to non-specific manifestations. Neuroimaging techniques are used in the diagnosis of NCC, however, due to the high cost of these methods and the advantages presented in the use of immunological tests, such as ease of performance and satisfactory results, immunoassays are commonly used to detect antibodies against Taenia sp. antigens. The aim of the present study was to produce, characterize and apply specific polyclonal immunoglobulin Y (IgY) anti-Taenia crassiceps extracted from egg yolk of hens immunized with T. crassiceps metacestodes.
Methods
Indirect enzyme-linked immunosorbent assay (ELISA), avidity ELISA, immunoblotting and indirect immunofluorescence tests were performed for characterization of IgY antibodies. Diagnostic performance was verified by ELISA for immune complex detection testing 90 serum samples.
Results
Values of sensitivity, specificity, positive and negative likelihood ratios (LR+/LR−) and area under the curve (AUC) were calculated and presented the following results: sensitivity 83.3%, specificity 96.7%, AUC 0.966, LR+ 25.0 and LR− 0.17.
Conclusions
Results of this pioneering and innovative study demonstrate that anti-T. crassiceps IgY antibodies present potential applicability and can be used as an efficient tool in human NCC serodiagnosis.
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