Allele specific PCR for a major marker of levamisole resistance in Haemonchus contortus.
Allele specific PCR for a major marker of levamisole resistance in Haemonchus contortus.
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DOI：
10.1016/j.ijpddr.2022.08.001
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发表时间：
2022-12
影响因子：
4
通讯作者：
Laing, Roz
中科院分区：
文献类型：
--
作者：
Antonopoulos, Alistair;Doyle, Stephen R.;Bartley, David J.;Morrison, Alison A.;Kaplan, Ray;Howell, Sue;Neveu, Cedric;Busin, Valentina;Devaney, Eileen;Laing, Roz
捻转血矛线虫（Haemonchus contortus）是一种食血寄生线虫，其感染小型反刍动物并在全球范围内在畜牧业中引起显著的动物健康问题和经济损失。治疗主要依靠广谱驱虫药，然而，对所有主要药物类别的耐药性已经建立或迅速出现。左旋咪唑（LEV）仍然是控制寄生虫的一个重要治疗选择，因为对LEV的耐药性不如对其他主要类别的驱虫药的耐药性普遍。LEV是一种乙酰胆碱受体（AChR）激动剂，当结合时，会导致蠕虫瘫痪。许多研究表明AChR亚单位ACR-8与LEV敏感性有关，特别是在一些耐药菌株中存在截短的acr-8转录物或acr-8基因座缺失。最近，acr-8中的一个单一的非同义SNP赋予丝氨酸到苏氨酸的取代（S168 T）被确定为与LEV抗性强烈相关。在这里，我们研究acr-8基因座的遗传变异在LEV敏感的MHco 3（伊势）和LEV抗性的MHco 18（UGA 2004）分离株之间的受控遗传杂交中的作用。扭曲使用单虫PCR检测，我们发现S168 T的存在与LEV抗性在亲本分离株和遗传杂交的F3后代中存活LEV处理强烈相关。我们开发并优化了等位基因特异性PCR检测S168 T，并验证了实验室分离株和现场样品，LEV耐药表型的测定。在LEV抗性田间种群中，高比例（>75%）的L3编码S168 T变异体，而该变异体在所研究的敏感菌株中不存在。这些数据进一步支持acr-8 S168 T在LEV抗性中的潜在作用，等位基因特异性PCR为建立LEV抗性的敏感分子诊断测试提供了重要的一步。
Haemonchus contortus is a haematophagous parasitic nematode that infects small ruminants and causes significant animal health concerns and economic losses within the livestock industry on a global scale. Treatment primarily depends on broad-spectrum anthelmintics, however, resistance is established or rapidly emerging against all major drug classes. Levamisole (LEV) remains an important treatment option for parasite control, as resistance to LEV is less prevalent than to members of other major classes of anthelmintics. LEV is an acetylcholine receptor (AChR) agonist that, when bound, results in paralysis of the worm. Numerous studies implicated the AChR sub-unit, ACR-8, in LEV sensitivity and in particular, the presence of a truncated acr-8 transcript or a deletion in the acr-8 locus in some resistant isolates. Recently, a single non-synonymous SNP in acr-8 conferring a serine-to-threonine substitution (S168T) was identified that was strongly associated with LEV resistance. Here, we investigate the role of genetic variation at the acr-8 locus in a controlled genetic cross between the LEV susceptible MHco3(ISE) and LEV resistant MHco18(UGA2004) isolates of H. contortus. Using single worm PCR assays, we found that the presence of S168T was strongly associated with LEV resistance in the parental isolates and F3 progeny of the genetic cross surviving LEV treatment. We developed and optimised an allele-specific PCR assay for the detection of S168T and validated the assay using laboratory isolates and field samples that were phenotyped for LEV resistance. In the LEV-resistant field population, a high proportion (>75%) of L3 encoded the S168T variant, whereas the variant was absent in the susceptible isolates studied. These data further support the potential role of acr-8 S168T in LEV resistance, with the allele-specific PCR providing an important step towards establishing a sensitive molecular diagnostic test for LEV resistance.
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影响因子：
4.5
作者：
Choi YJ;Bisset SA;Doyle SR;Hallsworth-Pepin K;Martin J;Grant WN;Mitreva M
通讯作者：
Mitreva M
影响因子：
3.3
作者：
Doyle SR;Laing R;Bartley DJ;Britton C;Chaudhry U;Gilleard JS;Holroyd N;Mable BK;Maitland K;Morrison AA;Tait A;Tracey A;Berriman M;Devaney E;Cotton JA;Sargison ND
通讯作者：
Sargison ND
影响因子：
4
作者：
Avramenko, Russell W.;Redman, Elizabeth M.;Gilleard, John S.
通讯作者：
Gilleard, John S.
影响因子：
3.8
作者：
Duguet, Thomas B.;Charvet, Claude L.;Beech, Robin N.
通讯作者：
Beech, Robin N.
影响因子：
4
作者：
Kopp, Steven R.;Coleman, Glen T.;Kotze, Andrew C.
通讯作者：
Kotze, Andrew C.