Optimal ELISA antigen for the diagnosis of Ascaris suum infection in humans

Yoshida, Ayako; Kikuchi, Taisei; Nakagaki, Shiori; Maruyama, Haruhiko

doi:10.1007/s00436-016-5239-3

Cited by 6 publications

(8 citation statements)

References 26 publications

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“…Only three individuals from this cohort had detectable Abs against T. canis (Tc-EcAD) and the same sera bound to somatic A. suum antigens (As-ExAD). In general, high levels of cross-reactivity between T. canis and A. suum has often been seen in ascarid infections due to the antigenic similarity between these two closely related species that cause human VLM syndrome [43,51]. Considering this and the fact that T. canis infection was confirmed by the highly specific Tc-ES-IB assay [42,51], these three patients were likely infected with T. canis and not with A. suum.…”

Section: Discussionmentioning

confidence: 97%

“…For detection of specific Abs to A. suum , the parasites were collected from the intestines of a naturally infected pig, and the soluble crude extract of a female worm (As-ExAD) was prepared as previously described [ 43 ] and used in a modified ELISA test [ 44 ]. Briefly, ELISA microplates (Nunc-Immuno TM Plate, Roskilde, Denmark) were coated with 1 μg/mL (100 ng/well) of As-ExAD in carbonate/bicarbonate buffer (0.05 M, pH 9.6).…”

Section: Methodsmentioning

confidence: 99%

“…The resulting optical densities (OD) were measured at 450 nm in a microplate ELISA reader (Filter-Max F5, Molecular Devices, San Jose, CA, USA). The samples were considered serologically positive if the OD values were higher than the cut-off point, i.e., the mean OD of nine negative control sera plus four times the standard deviation [43].…”

Section: Detection Of Specific Igg Antibodies To Toxocara Canis Ascamentioning

confidence: 99%

“…Larval ES antigens of T. canis (Tc-ES) were harvested from in vitro culture as described before [46]. The soluble whole-worm antigens (Tc-ExAD, As-ExAD) were prepared following the previously described procedure [43] and used in Western blot and ELISA [35] analysis. Toxocara canis larval ES products (Tc-ES) were used as a native antigen in both assays.…”

Section: Determination Of Antibody Levels To Ixodes Ricinus Salivary mentioning

confidence: 99%

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Infection with Toxocara canis Inhibits the Production of IgE Antibodies to α-Gal in Humans: Towards a Conceptual Framework of the Hygiene Hypothesis?

et al. 2020

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α-Gal syndrome (AGS) is a type of anaphylactic reaction to mammalian meat characterized by an immunoglobulin (Ig)E immune response to the oligosaccharide α-Gal (Galα1-3Galβ1-4GlcNAc-R). Tick bites seems to be a prerequisite for the onset of the allergic disease in humans, but the implication of non-tick parasites in α-Gal sensitization has also been deliberated. In the present study, we therefore evaluated the capacity of helminths (Toxocara canis, Ascaris suum, Schistosoma mansoni), protozoa (Toxoplasma gondii), and parasitic fungi (Aspergillus fumigatus) to induce an immune response to α-Gal. For this, different developmental stages of the infectious agents were tested for the presence of α-Gal. Next, the potential correlation between immune responses to α-Gal and the parasite infections was investigated by testing sera collected from patients with AGS and those infected with the parasites. Our results showed that S. mansoni and A. fumigatus produce the terminal α-Gal moieties, but they were not able to induce the production of specific antibodies. By contrast, T. canis, A. suum and T. gondii lack the α-Gal epitope. Furthermore, the patients with T. canis infection had significantly decreased anti-α-Gal IgE levels when compared to the healthy controls, suggesting the potential role of this nematode parasite in suppressing the allergic response to the glycan molecule. This rather intriguing observation is discussed in the context of the ‘hygiene hypothesis’. Taken together, our study provides new insights into the relationships between immune responses to α-Gal and parasitic infections. However, further investigations should be undertaken to identify T. canis components with potent immunomodulatory properties and to assess their potential to be used in immunotherapy and control of AGS.

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Section: Discussionmentioning

confidence: 97%

Section: Methodsmentioning

confidence: 99%

Section: Detection Of Specific Igg Antibodies To Toxocara Canis Ascamentioning

confidence: 99%

Section: Determination Of Antibody Levels To Ixodes Ricinus Salivary mentioning

confidence: 99%

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Infection with Toxocara canis Inhibits the Production of IgE Antibodies to α-Gal in Humans: Towards a Conceptual Framework of the Hygiene Hypothesis?

et al. 2020

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“…Other detection methods, including colorimetry, fluorescence, electrochemistry, chemiluminescence (CL) and electrochemiluminescence (ECL), have been performed on paper‐based immunodevices. Enzyme‐linked immunosorbent assay (ELISA) is a commonly used method. It combines the specificity of antibodies with the effective catalytic properties of enzymes for some specific analyte detection.…”

Section: Introductionmentioning

confidence: 99%

Three‐dimensional ring‐oven washing technique for a paper‐based immunodevice

et al. 2019

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Washing is a standard step for enzyme-linked immunosorbent assays (ELISA) performed on a paper-based chip, in which nonspecific-binding antibodies and antigens should be removed completely from the paper surface. In this study, a novel threedimensional (3D) washing strategy using a heating ring-oven was carried out on a paper-based chip. Compared with a plane washing mode by a ring-oven, this 3D washing strategy obtained a lower background, as gravity played an important role in the washing step. The paper-based chip was placed on a 3D plastic holder and the waste area was connected to a heating ring. Use of a heating waste area meant that the nonspecific-binding protein was continuously carried to the waste area through gravity and capillary action. The angle between the plastic holder and the ring plane was carefully selected. The effect of washing on different parts of the detection area was investigated by upconversion fluorescence and chemiluminescence (CL). This novel 3D washing strategy was performed for carcinoembryonic antigen detection through CL and a lower detection limit of 2 pg ml −1 was obtained. This approach provides an effective washing strategy to remove nonspecific-binding antibody from a paper-based immunodevice. K E Y W O R D Schemiluminescence, ELISA, nonspecific-binding, paper-based chip, three-dimensional washing

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