Haemonchus contortus is a parasitic haematophagous nematode that primarily affects small ruminants and causes significant economic loss to the global livestock industry. Treatment of haemonchosis typically relies on broad-spectrum anthelmintics, resistance to which is an important cause of treatment failure. Resistance to levamisole remains less widespread than to other major anthelmintic classes, prompting the need for more effective and accurate surveillance to maintain its efficacy. Loop-primer endonuclease cleavage loop-mediated isothermal amplification (LEC-LAMP) is a recently developed diagnostic method that facilitates multiplex target detection with single nucleotide polymorphism (SNP) specificity and portable onsite testing. In this study, we designed a new LEC-LAMP assay and applied it to detect the levamisole resistance marker S168T in H. contortus. We explored multiplexing probes for both the resistant S168T and the susceptible S168 alleles in a single-tube assay. We then included a generic probe to detect the acr-8 gene in the multiplex assay, which could facilitate the quantification of both resistance markers and overall genetic material from H. contortus in a single step. Our results showed promising application of these technologies, demonstrating a proof-of-concept assay which is amenable to detection of resistance alleles within the parasite population, with the potential for multiplex detection, and point-of-care application enabled by lateral flow end-point detection. However, further optimisation and validation is necessary.

捻转血矛线虫是一种寄生性吸血线虫，主要影响小型反刍动物，给全球畜牧业造成重大经济损失。血吸虫病的治疗通常依赖于广谱驱虫药，对其产生耐药性是治疗失败的重要原因。与其他主要驱虫药相比，左旋咪唑的耐药性仍然不那么普遍，因此需要更有效和准确的监测来维持其功效。环引物核酸内切酶裂解环介导的等温扩增 (LEC-LAMP) 是一种最近开发的诊断方法，可促进具有单核苷酸多态性 (SNP) 特异性的多重目标检测和便携式现场测试。在本研究中，我们设计了一种新的LEC-LAMP检测方法，并将其应用于检测H中的左旋咪唑耐药标记S168T。捻转。我们在单管测定中探索了抗性 S168T 和易感 S168 等位基因的多重探针。然后，我们在多重检测中加入了一个通用探针来检测 acr-8 基因，这可以促进抗性标记和总体遗传物质的量化H.一步内扭转。我们的结果显示了这些技术的应用前景，展示了一种概念验证测定法，该测定法适用于检测寄生虫群体内的抗性等位基因，具有多重检测的潜力，以及通过侧向流动末端实现的护理点应用点检测。但还需要进一步优化和验证。