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- Title
New Insights on Tools for Detecting β-Tubulin Polymorphisms in Trichuris trichiura Using rhAmp TM SNP Genotyping.
- Authors
Rivero, Julia; Cutillas, Cristina; Callejón, Rocío
- Abstract
Simple Summary: Soil-transmitted worm infections, usually treated with benzimidazoles, can develop resistance due to genetic variations in a specific gene called β-tubulin isotype 1. This study aimed to create a new, quick, and accurate way to identify these genetic variations. We designed a test to spot changes in certain parts of the β-tubulin gene of Trichuris trichiura, the worm causing the infection. By using two different primers, we could distinguish between different genetic types at specific positions in the gene. We tested our method on samples from captive primates and found it to be reliable. Additionally, we explored whether the β-tubulin gene could be useful as a marker in genetic studies. Our tests worked well when we used them on samples from the field. However, we did not find any of the expected genetic variations in the worms or eggs we examined. Instead, all the samples showed the same genetic type. Despite this, our analysis of the β-tubulin gene confirmed the close relationship between T. trichiura and a related Trichuris suis species, which suggests that this gene could be valuable for understanding their evolutionary history. Soil-transmitted helminth (STH) infections, commonly treated with benzimidazoles, are linked to resistance through single nucleotide polymorphisms (SNPs) at position 167, 198, or 200 in the β-tubulin isotype 1 gene. The aim of this study was to establish a novel genotyping assay characterized by its rapidity and specificity. This assay was designed to detect the presence of SNPs within the partial β-tubulin gene of Trichuris trichiura. This was achieved through the biallelic discrimination at codons 167, 198, and 200 by employing the competitive binding of two allele-specific forward primers. The specificity and reliability of this assay were subsequently confirmed using Trichuris samples isolated from captive primates. Furthermore, a molecular study was conducted to substantiate the utility of the β-tubulin gene as a molecular marker. The assays showed high sensitivity and specificity when applied to field samples. Nevertheless, none of the SNPs within the β-tubulin gene were detected in any of the adult worms or eggs from the analyzed populations. All specimens consistently displayed an SS genotype. The examination of the β-tubulin gene further validated the established close relationships between the T. trichiura clade and Trichuris suis clade. This reaffirms its utility as a marker for phylogenetic analysis.
- Subjects
TUBULINS; WHIPWORMS; WORM eggs; SINGLE nucleotide polymorphisms; GENETIC variation; GENETIC markers
- Publication
Animals (2076-2615), 2024, Vol 14, Issue 11, p1545
- ISSN
2076-2615
- Publication type
Article
- DOI
10.3390/ani14111545