Duplex Real-Time qPCR Assay Targeting β-Tubulin SNPs for Detection of Albendazole Resistance in Haemonchus contortus.
Chen X, Zhang L, Wang T, Zhang H, Wang R, Lu J, Gao W, Wang W, Liu C · Veterinary Journal · 22 March 2026
Duplex qPCR accurately detects albendazole resistance in H. contortus for targeted anthelmintic therapy.
This study describes the development and validation of a duplex real-time quantitative PCR (qPCR) assay for detecting albendazole resistance in Haemonchus contortus, a significant gastrointestinal parasitic nematode of ruminants. The assay targets single nucleotide polymorphisms at codons 198 (E198A) and 200 (F200Y) of the β-tubulin isotype I gene, which are the most frequently detected resistance-associated mutations in Chinese isolates. The optimized duplex qPCR demonstrated excellent analytical performance with detection limits of 1×10³ copies/μL for sensitive genotypes and 1×10 copies/μL for resistant genotypes, with intra-assay and inter-assay coefficients of variation below 0.55%. Clinical application to 102 samples identified 1 sensitive, 25 resistant, and 76 mixed infections. Sequencing validation of seven randomly selected samples confirmed qPCR accuracy. This rapid, sensitive, and specific molecular assay provides veterinary practitioners with an effective tool for accurate detection of albendazole resistance, enabling informed anthelmintic selection and surveillance of resistance trends in ruminant populations.
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