Immunoproteomic and Immunoinformatic Approaches Identify Sensitive Antigens for Diagnosing Anisakis pegreffii Infection.

IF 4 2区医学 Q2 CHEMISTRY, MEDICINAL ACS Infectious Diseases Pub Date : 2024-11-04 DOI:10.1021/acsinfecdis.4c00708

Xiaoxu Wang, Minhao Zeng, Guofeng Cheng

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Abstract

Anisakis 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 Anisakis infection. In the present study, we used immunoproteomic and immunoinformatic approaches to identify sensitive antigens for diagnosing Anisakis pegreffii 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 Gaussia luciferase immunoprecipitation assay (LIPS) was further developed upon the fusion of the proteins and Gaussia luciferase. The LIPS method indicated that A. pegreffii 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 A. pegreffii infection, particularly for the early stage.

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免疫蛋白组学和免疫形式学方法确定了诊断鹅口疮疥癣感染的敏感抗原。

疟原虫是一种食源性寄生虫，可伺机寄生于人体，导致急性腹部症状和过敏。除胃镜检查外，目前尚无其他诊断技术。因此，有必要确定特定的生物标志物，然后开发分子技术来诊断疟原虫感染。在本研究中，我们采用了免疫蛋白组学和免疫形式学方法来确定诊断虫弧菌感染的敏感抗原。根据免疫形式学结果，共鉴定出三种蛋白质，包括 Ani609（VDK51609）、Ani941（VDK75941）和 AniS13。然后，基于重组蛋白开发了间接 ELISA 方法，显示出与寄生虫可溶性蛋白相似的诊断能力。接着，在蛋白质与高斯荧光素酶融合的基础上，进一步开发了高斯荧光素酶免疫沉淀检测法（LIPS）。LIPS方法表明，大鼠感染A. pegreffii后最早可在1周内检测到，尤其是Ani941。总之，我们利用免疫蛋白组学和免疫形式学方法鉴定了新型抗原，然后开发了一种灵敏的方法来诊断 A. pegreffii 感染，尤其是早期感染。

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来源期刊

ACS Infectious Diseases CHEMISTRY, MEDICINALINFECTIOUS DISEASES&nb-INFECTIOUS DISEASES

CiteScore

9.70

自引率

3.80%

发文量

213

期刊介绍： ACS Infectious Diseases will be the first journal to highlight chemistry and its role in this multidisciplinary and collaborative research area. The journal will cover a diverse array of topics including, but not limited to: * Discovery and development of new antimicrobial agents — identified through target- or phenotypic-based approaches as well as compounds that induce synergy with antimicrobials. * Characterization and validation of drug target or pathways — use of single target and genome-wide knockdown and knockouts, biochemical studies, structural biology, new technologies to facilitate characterization and prioritization of potential drug targets. * Mechanism of drug resistance — fundamental research that advances our understanding of resistance; strategies to prevent resistance. * Mechanisms of action — use of genetic, metabolomic, and activity- and affinity-based protein profiling to elucidate the mechanism of action of clinical and experimental antimicrobial agents. * Host-pathogen interactions — tools for studying host-pathogen interactions, cellular biochemistry of hosts and pathogens, and molecular interactions of pathogens with host microbiota. * Small molecule vaccine adjuvants for infectious disease. * Viral and bacterial biochemistry and molecular biology.