Immunoproteomic and Immunoinformatic Approaches Identify Sensitive Antigens for Diagnosing <i>Anisakis pegreffii</i> Infection

<i>Anisakis</i> are foodborne parasites that opportunistically parasitize humans, leading to acute abdominal symptoms and allergies. Besides gastroscopy, no other diagnostic technique is available. Consequently, it is necessary to identify specific biomarkers and then develop molecular techniques for diagnosing <i>Anisakis</i> infection. In the present study, we used immunoproteomic and immunoinformatic approaches to identify sensitive antigens for diagnosing <i>Anisakis pegreffii</i> infection. A total of three proteins, including Ani609 (VDK51609), Ani941 (VDK75941), and AniS13, were identified based on immunoinformatic results. Then, the indirect ELISA method was developed based on the recombinant proteins, showing a similar diagnostic capability to that of parasitic soluble proteins. Next, a <i>Gaussia</i> luciferase immunoprecipitation assay (LIPS) was further developed upon the fusion of the proteins and <i>Gaussia</i> luciferase. The LIPS method indicated that <i>A. pegreffii</i> infection could be detected in rats as early as 1 week post infection, especially for Ani941. Overall, we identified the novel antigens using immunoproteomic and immunoinformatic approaches and then developed a sensitive method for diagnosing <i>A. pegreffii</i> infection, particularly for the early stage.