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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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Development of nucleic acid lateral flow immunoassay for molecular detection of Entamoeba moshkovskii and Entamoeba dispar in stool samples
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Journal Article
Development of nucleic acid lateral flow immunoassay for molecular detection of Entamoeba moshkovskii and Entamoeba dispar in stool samples
2024
Overview
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 fg of
E. moshkovskii
target DNA (982 parasites or 196 parasites/microliter), and 487.5 fg 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.
Publisher
Nature Publishing Group UK,Nature Publishing Group,Nature Portfolio
Subject
