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巴西人群中毛首鞭形线虫β-微管蛋白基因中苯并咪唑抗性相关单核苷酸多态性的检测

Detection of Benzimidazole Resistance-Associated Single-Nucleotide Polymorphisms in the Beta-Tubulin Gene in Trichuris trichiura from Brazilian Populations.

作者信息

Mendes de Oliveira Valéria Nayara Gomes, Zuccherato Luciana Werneck, Dos Santos Talita Rodrigues, Rabelo Élida Mara Leite, Furtado Luis Fernando Viana

机构信息

Universidade Federal de Minas Gerais, Instituto de Ciências Biológicas, Belo Horizonte, Minas Gerais, Brazil.

Universidade do Estado de Minas Gerais, Unidade Passos, Minas Gerais, Brazil.

出版信息

Am J Trop Med Hyg. 2022 Jul 25;107(3):640-8. doi: 10.4269/ajtmh.22-0157.

Abstract

Preventive chemotherapy is recommended by the WHO as the main strategy for controlling infections caused by nematodes in humans, aiming to eliminate the morbidity associated with these infections. This strategy consists of routine periodic administration of benzimidazoles, among other drugs. Although these drugs decrease the intensity of infections, they have the potential to exert selection pressure for genotypes bearing mutations associated with drug resistance, which may result in the establishment of resistant worm populations. There is evidence in the literature of resistance to these drugs in nematodes that infect humans, including in the species Trichuris trichiura. Single-nucleotide polymorphisms (SNPs) in the beta-tubulin gene located at codons 167, 198, and 200 are associated with the mechanism of resistance to benzimidazoles in nematodes. Here, we standardized a molecular technique based on an amplification refractory mutation system-polymerase chain reaction (ARMS-PCR) to analyze codons 167, 198, and 200 of T. trichiura. The ARMS-PCR methodology was successfully established to evaluate the codons of interest. A total of 420 samples of individual eggs were analyzed from populations obtained from five Brazilian states. A mutation in codon 198 was observed at a frequency of 4.8% (20/420), while for the other two codons, no polymorphism was observed. This is the first report of the presence of this mutation in populations of T. trichiura in Brazil. This fact and the emergence of the problem already observed in other species reinforces the need for regular monitoring of SNPs related to benzimidazole resistance using techniques that are highly sensitive and specific.

摘要

世界卫生组织推荐预防性化疗作为控制人类线虫感染的主要策略,旨在消除与这些感染相关的发病率。该策略包括定期常规施用苯并咪唑等药物。尽管这些药物可降低感染强度,但它们有可能对携带与耐药性相关突变的基因型施加选择压力,这可能导致耐药虫种群的形成。文献中有证据表明,包括毛首鞭形线虫在内的感染人类的线虫对这些药物存在耐药性。位于密码子167、198和200处的β-微管蛋白基因中的单核苷酸多态性(SNP)与线虫对苯并咪唑的耐药机制有关。在此,我们基于扩增阻滞突变系统-聚合酶链反应(ARMS-PCR)标准化了一种分子技术,以分析毛首鞭形线虫的密码子167、198和200。成功建立了ARMS-PCR方法来评估感兴趣的密码子。对来自巴西五个州的种群的420个单个虫卵样本进行了分析。观察到密码子198处的突变频率为4.8%(20/420),而对于其他两个密码子,未观察到多态性。这是巴西毛首鞭形线虫种群中存在这种突变的首次报告。这一事实以及在其他物种中已经观察到的问题的出现,强化了使用高度敏感和特异的技术定期监测与苯并咪唑耐药性相关的SNP的必要性。

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