New Insights on Tools for Detecting β-Tubulin Polymorphisms in Trichuris trichiura Using rhAmp^TM SNP Genotyping

Area of Science:

• Molecular parasitology
• Antiparasitic drug resistance
• Genetic analysis of helminths

Background:

• Soil-transmitted helminth (STH) infections are prevalent globally.
• Benzimidazole drugs are primary treatments for STH.
• Drug resistance, often due to SNPs in the β-tubulin gene, is a growing concern.

Purpose of the Study:

• To develop a rapid and specific genotyping assay for detecting SNPs in the β-tubulin gene of Trichuris trichiura.
• To investigate the prevalence of resistance-associated SNPs in T. trichiura populations.
• To confirm the utility of the β-tubulin gene as a molecular marker for phylogenetic studies.

Main Methods:

• Developed a novel assay using competitive binding of allele-specific primers for biallelic discrimination at codons 167, 198, and 200 of the β-tubulin gene.
• Validated the assay's specificity and reliability using Trichuris samples from captive primates.
• Applied the assay to field samples of adult worms and eggs.
• Conducted phylogenetic analysis using the β-tubulin gene.

Main Results:

• The developed assay demonstrated high sensitivity and specificity.
• No single nucleotide polymorphisms (SNPs) associated with benzimidazole resistance were detected in any of the analyzed T. trichiura adult worms or eggs.
• All specimens consistently showed an SS genotype.
• Phylogenetic analysis confirmed close evolutionary relationships between T. trichiura and Trichuris suis.

Conclusions:

• The novel genotyping assay is effective for detecting resistance SNPs in T. trichiura.
• The studied T. trichiura populations do not exhibit detectable benzimidazole resistance via SNPs in the β-tubulin gene.
• The β-tubulin gene is a reliable molecular marker for T. trichiura phylogenetic studies.