内蒙古地区捻转血矛线虫阿苯达唑耐药种群Ⅰ型β微管蛋白基因单核苷酸多态性调查

Investigation on SNPs of Isotype-Ⅰβ-tubulin Gene of Haemonchus contortus Populations of Albendazole Resistance in Inner Mongolia

为了解内蒙古地区捻转血矛线虫阿苯达唑耐药虫株Ⅰ型β微管蛋白基因单核苷酸多态性的情况,对来自内蒙古乌审旗、察右后旗和科右前旗3个地区各19条捻转血矛线虫雄虫进行ITS-2基因特异性PCR鉴定为捻转血矛线虫后,扩增出Ⅰ型β微管蛋白基因,测序后对耐药相关167、198和200位点突变情况进行调查分析。结果表明,在这3个地区均未发现167位点突变,主要是198和200位点发生突变,总体上看198位点突变频率高于200位点。分析这57条雄虫基因序列发现4种基因型,分别是198纯合耐药型及200纯合敏感型、198纯合敏感型及200纯合耐药型、198纯合敏感型及200杂合耐药型和198及200均杂合耐药型。而耐药基因型频率乌审旗为100%,察右后旗68.42%,科右前旗94.74%,这与粪便虫卵减少试验结果一致。研究结果为捻转血矛线虫病的预防和治疗提供一个新思路,可有效避免耐药性或者减缓耐药性的产生,减少畜牧养殖行业的经济损失。

To understand the situation of single nucleotide polymorphisms(SNP)of isotype-Ⅰβ-tubulin gene of Haemonchus contortus albendazole-resistant strains in Inner Mongolia,the ITS-2 gene of 19 male species of Haemonchus contortus from Uxin Qi,Qahar Youyi Houqi and Horqin Youyi Qianqi in Inner Mongolia was identified by specific PCR as Haemonchus contortus,and isotype-Ⅰβ-tubulin gene was amplified,sequenced and analyzed for investigation of benzimidazole-related SNPs at loci 167,198 and 200.The results showed that F167Y was not found in these three regions,and E198A and F200Y were mainly found.Generally speaking,the frequency of E198A was higher than that of F200Y.Analysis of these 57 male gene sequences revealed four genotypes,which were rr-198 and SS-200,SS-198 and rr-200,SS-198 and rS-200,and rS-198 and rS-200.The genotype frequency of drug resistance was 100%in Uxin Qi,68.42%in Qahar Youyi Houqi and 94.74%in Horqin Youyi Qianqi,which was consistent with FECRT.The results provided a new idea for the prevention and treatment of Haemonchus contortus,which can effectively avoid or slow down the development of drug resistance,and reduce economic losses in the livestock breeding industry.