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A Molecular Diagnostic Tool to Replace Larval Culture in Conventional Faecal Egg Count Reduction Testing in Sheep

机译:替代绵羊常规粪便卵数检测中幼虫培养的分子诊断工具

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摘要

The accurate diagnosis of parasitic nematode infections in livestock (including sheep and goats) is central to their effective control and the detection of the anthelmintic resistance. Traditionally, the faecal egg count reduction test (FECRT), combined with the technique of larval culture (LC), has been used widely to assess drug-susceptibility/resistance in strongylid nematodes. However, this approach suffers from a lack of specificity, sensitivity and reliability, and is time-consuming and costly to conduct. Here, we critically assessed a specific PCR assay to support FECRT, in a well-controlled experiment on sheep with naturally acquired strongylid infections known to be resistant to benzimidazoles. We showed that the PCR results were in close agreement with those of total worm count (TWC), but not of LC. Importantly, albendazole resistance detected by PCR-coupled FECRT was unequivocally linked to Teladorsagia circumcincta and, to lesser extent, Trichostrongylus colubriformis, a result that was not achievable by LC. The key findings from this study demonstrate that our PCR-coupled FECRT approach has major merit for supporting anthelmintic resistance in nematode populations. The findings also show clearly that our PCR assay can be used as an alternative to LC, and is more time-efficient and less laborious, which has important practical implications for the effective management and control strongylid nematodes of sheep.
机译:准确诊断家畜(包括绵羊和山羊)的寄生线虫感染是对其进行有效控制和检测驱虫药耐药性的关键。传统上,粪便卵数减少测试(FECRT)与幼虫培养(LC)技术相结合已广泛用于评估强线虫线虫的药物敏感性/耐药性。但是,这种方法缺乏特异性,敏感性和可靠性,并且耗时且昂贵。在这里,我们在对绵羊进行自然控制的实验中,严格评估了一种支持FECRT的特异性PCR检测方法,该绵羊患有自然获得的强甲虫感染,已知对苯并咪唑具有抗性。我们显示PCR结果与蠕虫总数(TWC)的结果非常一致,但LC结果却不一致。重要的是,通过PCR耦合的FECRT检测到的阿苯达唑耐药性明确地与Teladorsagia circumcincta以及在较小程度上与cochoriformis的Trichostrongylus连锁,这是LC无法实现的。这项研究的主要发现表明,我们的PCR偶联FECRT方法具有支持线虫种群中驱虫抗药性的主要优点。研究结果还清楚地表明,我们的PCR测定法可以替代LC,而且效率更高,更省时,更省力,这对有效管理和控制绵羊线虫的线虫具有重要的实际意义。

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