A novel cross-priming amplification technique combined with lateral flow strips for rapid and visual detection of zoonotic Toxoplasma gondii

IF 2 2区农林科学 Q2 PARASITOLOGY

Veterinary parasitology Pub Date : 2025-02-01 DOI:10.1016/j.vetpar.2025.110402

Yao Liang , Yuan-Hui He , Shu-Feng Yang , Shi-Chen Xie , Yi-Han Lv , Wei Cong , Hany M. Elsheikha , Xing-Quan Zhu

{"title":"A novel cross-priming amplification technique combined with lateral flow strips for rapid and visual detection of zoonotic Toxoplasma gondii","authors":"Yao Liang , Yuan-Hui He , Shu-Feng Yang , Shi-Chen Xie , Yi-Han Lv , Wei Cong , Hany M. Elsheikha , Xing-Quan Zhu","doi":"10.1016/j.vetpar.2025.110402","DOIUrl":null,"url":null,"abstract":"<div><div><em>Toxoplasma gondii</em>, an obligate intracellular protozoan, infects almost all warm-blooded animals and humans, with felines serving as its sole definitive hosts. Cats release <em>T. gondii</em> oocysts into the environment through feces, contributing to environmental contamination that can lead to toxoplasmosis in humans upon exposure through ingestion of contaminated food, water, or soil. Effective detection of <em>T. gondii</em> in environmental samples is essential for protecting public health and preventing disease transmission. In the present study, we developed a cross-priming amplification (CPA) assay coupled with lateral flow immunoassay strips for the rapid and visual detection of <em>T. gondii</em> in environmental samples. CPA offers simplicity and eliminates the need for complex laboratory equipment. The assay demonstrated high specificity, accurately identifying nine genotypes of <em>T. gondii</em> without cross-reacting with 11 related parasites. Sensitivity testing revealed a detection limit of 1 × 10² copies/μL at the molecular level (plasmid) and 10 oocysts in real-world environmental samples. Furthermore, CPA effectively detected <em>T. gondii</em> in diverse environmental samples, including soil, water, and cat feces, with results consistent with known infection rates. These findings underscore CPA′s potential as a reliable, rapid, and accessible tool for detecting <em>T. gondii</em> in environmental settings, contributing to improved public health surveillance and disease prevention.</div></div>","PeriodicalId":23716,"journal":{"name":"Veterinary parasitology","volume":"334 ","pages":"Article 110402"},"PeriodicalIF":2.0000,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Veterinary parasitology","FirstCategoryId":"97","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S0304401725000135","RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"PARASITOLOGY","Score":null,"Total":0}

引用次数: 0

Abstract

Toxoplasma gondii, an obligate intracellular protozoan, infects almost all warm-blooded animals and humans, with felines serving as its sole definitive hosts. Cats release T. gondii oocysts into the environment through feces, contributing to environmental contamination that can lead to toxoplasmosis in humans upon exposure through ingestion of contaminated food, water, or soil. Effective detection of T. gondii in environmental samples is essential for protecting public health and preventing disease transmission. In the present study, we developed a cross-priming amplification (CPA) assay coupled with lateral flow immunoassay strips for the rapid and visual detection of T. gondii in environmental samples. CPA offers simplicity and eliminates the need for complex laboratory equipment. The assay demonstrated high specificity, accurately identifying nine genotypes of T. gondii without cross-reacting with 11 related parasites. Sensitivity testing revealed a detection limit of 1 × 10² copies/μL at the molecular level (plasmid) and 10 oocysts in real-world environmental samples. Furthermore, CPA effectively detected T. gondii in diverse environmental samples, including soil, water, and cat feces, with results consistent with known infection rates. These findings underscore CPA′s potential as a reliable, rapid, and accessible tool for detecting T. gondii in environmental settings, contributing to improved public health surveillance and disease prevention.

查看原文本刊更多论文

一种新的交叉引物扩增技术结合横向流动条带快速和视觉检测人畜共患弓形虫。

刚地弓形虫是一种专性细胞内原生动物，几乎感染所有温血动物和人类，猫科动物是其唯一的最终宿主。猫通过粪便将弓形虫卵囊释放到环境中，造成环境污染，人类通过摄入受污染的食物、水或土壤而感染弓形虫病。有效检测环境样本中的弓形虫对保护公众健康和预防疾病传播至关重要。在本研究中，我们建立了一种交叉引物扩增（CPA）方法，结合横向流动免疫测定条，用于快速和视觉检测环境样品中的弓形虫。CPA提供简单性，消除了对复杂实验室设备的需求。该方法具有较高的特异性，可准确鉴定出9种基因型的弓形虫，而不会与11种相关寄生虫发生交叉反应。灵敏度测试结果表明，在分子水平（质粒）检测限为1 × 10²copies/μL，在实际环境样品中检测限为10个卵囊。此外，CPA有效地检测了土壤、水和猫粪便等不同环境样本中的弓形虫，结果与已知感染率一致。这些发现强调了CPA作为在环境环境中检测弓形虫的可靠、快速和可获得的工具的潜力，有助于改善公共卫生监测和疾病预防。

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

Veterinary parasitology 农林科学-寄生虫学

CiteScore

5.30

自引率

7.70%

发文量

126

审稿时长

36 days

期刊介绍： The journal Veterinary Parasitology has an open access mirror journal,Veterinary Parasitology: X, sharing the same aims and scope, editorial team, submission system and rigorous peer review. This journal is concerned with those aspects of helminthology, protozoology and entomology which are of interest to animal health investigators, veterinary practitioners and others with a special interest in parasitology. Papers of the highest quality dealing with all aspects of disease prevention, pathology, treatment, epidemiology, and control of parasites in all domesticated animals, fall within the scope of the journal. Papers of geographically limited (local) interest which are not of interest to an international audience will not be accepted. Authors who submit papers based on local data will need to indicate why their paper is relevant to a broader readership. Parasitological studies on laboratory animals fall within the scope of the journal only if they provide a reasonably close model of a disease of domestic animals. Additionally the journal will consider papers relating to wildlife species where they may act as disease reservoirs to domestic animals, or as a zoonotic reservoir. Case studies considered to be unique or of specific interest to the journal, will also be considered on occasions at the Editors'' discretion. Papers dealing exclusively with the taxonomy of parasites do not fall within the scope of the journal.