Validation of an optimized in-house enzyme-linked immunosorbent assay for enhanced detection of Trichinella spp. exposure in swine

IF 3.1 Q2 PARASITOLOGY

Food and Waterborne Parasitology Pub Date : 2026-03-01 Epub Date: 2026-02-19 DOI:10.1016/j.fawpar.2026.e00322

Vladislav A. Lobanov, Kelly A. Konecsni, W. Brad Scandrett

{"title":"Validation of an optimized in-house enzyme-linked immunosorbent assay for enhanced detection of Trichinella spp. exposure in swine","authors":"Vladislav A. Lobanov, Kelly A. Konecsni, W. Brad Scandrett","doi":"10.1016/j.fawpar.2026.e00322","DOIUrl":null,"url":null,"abstract":"<div><div>The International Commission on Trichinellosis (ICT) and the World Organization for Animal Health (WOAH) recommend the use of an indirect enzyme-linked immunosorbent assay (ELISA) that utilizes excretory-secretory (E-S) antigens (ESA) of Trichinella spiralis for surveillance and epidemiological studies in pigs and wild boars. Our efforts to optimize and standardize ESA production and ELISA protocols resulted in improved diagnostic performance of an in-house E-S ELISA. To validate the optimized assay, we compared its performance with that of a commercial E-S ELISA kit using sera from a representative set of commercial Canadian pigs (presumably Trichinella-free) and pigs experimentally infected with Trichinella spp. Both assays correctly identified the positive and negative sera, yielding 100% diagnostic sensitivity and specificity. However, in-house E-S ELISA exhibited a higher resolving power, as evidenced by the markedly better separation of normalized absorbance values of positive sera from those of samples from the negative pig population. Furthermore, significantly higher serial dilutions of sera from pigs experimentally infected with T. spiralis, T. pseudospiralis, T. britovi and T. nativa tested positive by the in-house E-S ELISA, demonstrating a higher analytical sensitivity of this assay. We continued testing sera from Canadian commercial pigs using the in-house assay to obtain a more accurate estimate of its diagnostic specificity. A total of 6345 animals have been tested, with only 11 samples showing test values above the cut-off. Ten of these sera tested negative by confirmatory western blot (WB). Therefore, the diagnostic specificity of in-house E-S ELISA alone and in combination with WB testing was 99.84% and 99.98%, respectively. WB detected seroconversion earlier than the optimized E-S ELISA in five of 15 pigs experimentally infected with various low doses of T. spiralis. The results of this study support the use of the optimized E-S ELISA and confirmatory WB for epidemiological surveys to monitor exposure to Trichinella spp. in swine.</div></div>","PeriodicalId":37941,"journal":{"name":"Food and Waterborne Parasitology","volume":"42 ","pages":"Article e00322"},"PeriodicalIF":3.1000,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Food and Waterborne Parasitology","FirstCategoryId":"1085","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S2405676626000089","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"2026/2/19 0:00:00","PubModel":"Epub","JCR":"Q2","JCRName":"PARASITOLOGY","Score":null,"Total":0}

引用次数: 0

Abstract

The International Commission on Trichinellosis (ICT) and the World Organization for Animal Health (WOAH) recommend the use of an indirect enzyme-linked immunosorbent assay (ELISA) that utilizes excretory-secretory (E-S) antigens (ESA) of Trichinella spiralis for surveillance and epidemiological studies in pigs and wild boars. Our efforts to optimize and standardize ESA production and ELISA protocols resulted in improved diagnostic performance of an in-house E-S ELISA. To validate the optimized assay, we compared its performance with that of a commercial E-S ELISA kit using sera from a representative set of commercial Canadian pigs (presumably Trichinella-free) and pigs experimentally infected with Trichinella spp. Both assays correctly identified the positive and negative sera, yielding 100% diagnostic sensitivity and specificity. However, in-house E-S ELISA exhibited a higher resolving power, as evidenced by the markedly better separation of normalized absorbance values of positive sera from those of samples from the negative pig population. Furthermore, significantly higher serial dilutions of sera from pigs experimentally infected with T. spiralis, T. pseudospiralis, T. britovi and T. nativa tested positive by the in-house E-S ELISA, demonstrating a higher analytical sensitivity of this assay. We continued testing sera from Canadian commercial pigs using the in-house assay to obtain a more accurate estimate of its diagnostic specificity. A total of 6345 animals have been tested, with only 11 samples showing test values above the cut-off. Ten of these sera tested negative by confirmatory western blot (WB). Therefore, the diagnostic specificity of in-house E-S ELISA alone and in combination with WB testing was 99.84% and 99.98%, respectively. WB detected seroconversion earlier than the optimized E-S ELISA in five of 15 pigs experimentally infected with various low doses of T. spiralis. The results of this study support the use of the optimized E-S ELISA and confirmatory WB for epidemiological surveys to monitor exposure to Trichinella spp. in swine.

查看原文本刊更多论文

一种优化的内部酶联免疫吸附试验的验证，用于增强猪旋毛虫暴露的检测

国际旋毛虫病委员会（ICT）和世界动物卫生组织（WOAH）建议使用间接酶联免疫吸附试验（ELISA），利用旋毛虫的排泄-分泌（E-S）抗原（ESA）对猪和野猪进行监测和流行病学研究。我们努力优化和标准化ESA生产和ELISA协议，从而提高了内部E-S ELISA的诊断性能。为了验证优化后的检测方法，我们将其与商用E-S ELISA试剂盒的性能进行了比较，该试剂盒使用了一组具有代表性的加拿大商用猪（可能不含旋毛虫）和实验感染旋毛虫的猪的血清，两种检测方法都能正确识别阳性和阴性血清，诊断灵敏度和特异性均为100%。然而，内部E-S ELISA具有更高的分辨能力，阳性血清与阴性猪群样品的归一化吸光度值的分离效果明显更好。此外，E-S酶联免疫吸附试验（ELISA）对实验感染螺旋体、假螺旋体、布氏体和原生螺旋体的猪血清的连续稀释检测结果呈阳性，表明该试验具有较高的分析敏感性。我们继续使用内部测定法检测加拿大商品猪的血清，以获得更准确的诊断特异性估计。总共对6345只动物进行了测试，只有11只样本的测试值高于临界值。其中10例血清经确认性免疫印迹（WB）检测为阴性。因此，内部E-S ELISA单独和联合WB检测的诊断特异性分别为99.84%和99.98%。在实验感染不同低剂量螺旋体的15头猪中，WB检测到的血清转化比优化后的E-S ELISA检测到的早。本研究结果支持采用优化后的E-S酶联免疫吸附试验和验证性WB进行流行病学调查，以监测猪对旋毛虫的暴露。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

求助全文

约1分钟内获得全文求助全文

来源期刊

Food and Waterborne Parasitology Immunology and Microbiology-Parasitology

CiteScore

5.10

自引率

4.00%

发文量

审稿时长

13 weeks

期刊介绍： Food and Waterborne Parasitology publishes high quality papers containing original research findings, investigative reports, and scientific proceedings on parasites which are transmitted to humans via the consumption of food or water. The relevant parasites include protozoa, nematodes, cestodes and trematodes which are transmitted by food or water and capable of infecting humans. Pertinent food includes products of animal or plant origin which are domestic or wild, and consumed by humans. Animals and plants from both terrestrial and aquatic sources are included, as well as studies related to potable and other types of water which serve to harbor, perpetuate or disseminate food and waterborne parasites. Studies dealing with prevalence, transmission, epidemiology, risk assessment and mitigation, including control measures and test methodologies for parasites in food and water are of particular interest. Evidence of the emergence of such parasites and interactions among domestic animals, wildlife and humans are of interest. The impact of parasites on the health and welfare of humans is viewed as very important and within scope of the journal. Manuscripts with scientifically generated information on associations between food and waterborne parasitic diseases and lifestyle, culture and economies are also welcome. Studies involving animal experiments must meet the International Guiding Principles for Biomedical Research Involving Animals as issued by the Council for International Organizations of Medical Sciences.