Novel assay for the detection and monitoring of levamisole resistance in Haemonchus contortus
Novel assay for the detection and monitoring of levamisole resistance in Haemonchus contortus
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DOI：
10.1016/j.ijpara.2013.12.004
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发表时间：
2014-03-01
影响因子：
4
通讯作者：
Prichard, Roger K.
中科院分区：
文献类型：
--
作者：
Barrere, Virginie;Beech, Robin N.;Prichard, Roger K.
左旋咪唑敏感的乙酰胆碱受体最近已被描述在寄生线虫Hoemonchus contortus。五聚体受体由不同的亚基蛋白产生，其中之一是Hco-ACR-8。Hco-acr-8b是一个截短的转录本，在6个左旋咪唑抗性H.结果表明，Hco-acr-8b在4株左旋咪唑敏感菌株中均缺失，提示Hco-acr-8b可能是左旋咪唑抗性的潜在标记。Hco-acr-8b转录物含有外显子1和2，并终止于内含子2内的347 bp。在这项工作中，我们研究了Hco-acr-8基因的基因组DNA序列，在一个区域，包括外显子2和外显子3，从广泛的左旋咪唑敏感和耐药的H。扭体线虫分离株从个体和寄生虫库中分析了可能参与产生第二内含子内截短剪接变体的序列。我们发现了一个插入/缺失（插入/缺失）63 bp位于下游的剪接受体位点的第三外显子的选择。插入缺失的序列，当存在时，在检查的12个分离株中是相似的。该插入缺失的存在或不存在与左旋咪唑耐药状态在统计学上相关（卡方检验）。indel的缺失与Hco-acr-8b转录本的表达之间也存在相关性。我们相信这是第一个报告的一个推定的DNA标记左旋咪唑耐药检测。利用这一新的知识，我们已经开发了一种新的基于DNA的检测和监测动物寄生线虫左旋咪唑耐药性的方法。(C)2014年澳大利亚寄生虫学会爱思唯尔有限公司版权所有
A levamisole-sensitive acetylcholine receptor has recently been described in the parasitic nematode Hoemonchus contortus. The pentameric receptor is produced from different subunit proteins, one of which is Hco-ACR-8. A truncated transcript, Hco-acr-8b, has been identified in six levamisole-resistant H. contortus isolates and was found to be absent in four levamisole-susceptible isolates, indicating Hco-acr-8b could be a potential marker for levamisole resistance. The Hco-acr-8b transcript contains exons 1 and 2 and terminates with 347 bp from within the intron 2. In this work, we investigated genomic DNA sequences of the Hco-acr-8 gene, in a region including exon 2 and exon 3, from a wide range of levamisole-susceptible and resistant H. contortus isolates. Sequences potentially involved in generating the truncated splice variant within the second intron were analysed from individuals and pools of parasites. We found an insertion/deletion (indel) of 63 bp located just downstream from the splice acceptor site for the alternative third exon. The sequence of the indel, when present, was similar in the 12 isolates examined. The presence or absence of this indel was statistically (Chi(2) test) correlated with levamisole resistance status. A correlation was also demonstrated between the absence of the indel and the expression of the Hco-acr-8b transcript. We believe this is the first report of a putative DNA marker for levamisole resistance detection. Using this new knowledge, we have developed a novel DNA-based assay for the detection and monitoring of levamisole resistance in parasitic nematodes of animals. (C) 2014 Australian Society for Parasitology Inc. Published by Elsevier Ltd. All rights reserved