PDF(Pubmed)
Abstract：
Soil-transmitted helminth (STH) infections, commonly treated with benzimidazoles, are linked to resistance through single nucleotide polymorphisms (SNPs) at position 167, 198, or 200 in the β-tubulin isotype 1 gene. The aim of this study was to establish a novel genotyping assay characterized by its rapidity and specificity. This assay was designed to detect the presence of SNPs within the partial β-tubulin gene of Trichuris trichiura. This was achieved through the biallelic discrimination at codons 167, 198, and 200 by employing the competitive binding of two allele-specific forward primers. The specificity and reliability of this assay were subsequently confirmed using Trichuris samples isolated from captive primates. Furthermore, a molecular study was conducted to substantiate the utility of the β-tubulin gene as a molecular marker. The assays showed high sensitivity and specificity when applied to field samples. Nevertheless, none of the SNPs within the β-tubulin gene were detected in any of the adult worms or eggs from the analyzed populations. All specimens consistently displayed an SS genotype. The examination of the β-tubulin gene further validated the established close relationships between the T. trichiura clade and Trichuris suis clade. This reaffirms its utility as a marker for phylogenetic analysis.
摘要：
土壤传播的蠕虫（STH）感染，通常用苯并咪唑治疗，通过β-微管蛋白同种型1基因中第167、198或200位的单核苷酸多态性（SNP）与抗性相关。这项研究的目的是建立一种新颖的基因分型测定法，其特征在于其快速性和特异性。设计该测定以检测Trichuris的部分β-微管蛋白基因内SNP的存在。这是通过使用两个等位基因特异性正向引物的竞争性结合在密码子167、198和200处的双等位基因区分来实现的。该测定的特异性和可靠性随后使用从圈养的灵长类动物分离的鞭虫样品进行确认。此外，进行了一项分子研究，以证实β-微管蛋白基因作为分子标记的实用性。当应用于现场样品时，该测定法显示出高灵敏度和特异性。然而，在来自分析种群的任何成虫或卵中都未检测到β-微管蛋白基因内的SNP。所有标本一致显示SS基因型。对β-微管蛋白基因的检查进一步验证了T.trichiura进化枝和猪Trichuris进化枝之间已建立的密切关系。这重申了其作为系统发育分析标记的实用性。
