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Development of nucleic acid lateral flow immunoassay for molecular detection of Entamoeba moshkovskii and Entamoeba dispar in stool samples
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Document Title
Development of nucleic acid lateral flow immunoassay for molecular detection of Entamoeba moshkovskii and Entamoeba dispar in stool samples
Author
Hutagalung S.V. Rattaprasert P. Promptmas C. Moonsom S. Yongkiettrakul S. Thima K. Chavalitshewinkoon-Petmitr P.
Affiliations
Department of Protozoology Faculty of Tropical Medicine Mahidol University Bangkok 10400 Thailand; Department of Biomedical Engineering Faculty of Engineering Mahidol University Nakhon Pathom 73170 Thailand; National Center for Genetic Engineering and Biotechnology Pathumthani 12120 Thailand
Type
Article
Source Title
Scientific Reports
ISSN
20452322
Year
2024
Volume
14
Issue
1
Open Access
All Open Access Gold Green
Publisher
Nature Research
DOI
10.1038/s41598-024-57332-3
Abstract
Entamoeba moshkovskii recently known as a possible pathogenic amoeba and the non-pathogenic Entamoeba dispar are morphologically indistinguishable by microscopy. Although PCR was used for differential diagnosis gel electrophoresis is labor-intensive time-consuming and exposed to hazardous elements. In this study nucleic acid lateral flow immunoassay (NALFIA) was developed to detect E. moshkovskii and E. dispar by post-PCR amplicon analysis. E. moshkovskii primers were labeled with digoxigenin and biotin whereas primers of E. dispar were lebeled with FITC and digoxigenin. The gold nanoparticles were labeled with antibodies corresponding to particular labeling. Based on the established assay NALFIA could detect as low as 975爁g of E. moshkovskii target DNA (982 parasites or 196 parasites/microliter) and 487.5爁g of E. dispar target DNA (444 parasites or 89 parasites/microliter) without cross-reactivity to other tested intestinal organisms. After testing 91 stool samples NALFIA was able to detect seven E. moshkovskii (87.5% sensitivity and 100% specificity) and eight E. dispar samples (66.7% sensitivity and 100% specificity) compared to real-time PCR. Interestingly it detected three mixed infections as real-time PCR. Therefore it can be a rapid safe and effective method for the detection of the emerging pathogens E. moshkovskii and E. dispar in stool samples. ? The Author(s) 2024.
License
CC BY
Rights
Authors
Publication Source
WOS