Screening And Detection Methods Of Albendazole Resistance Related Genes In Haemonchus Contortus

Haemonchus contortus is a common parasitic nematode in ruminants such as cattle and sheep.To screen albendazole resistance molecular marker genes of Haemonchus contortus,and established a drug resistance detection method of Haemonchus contortus for clinical guidance.In this study,the transcriptome data of albendazole sensitive and drug-resistant strains of Haemonchus contortus were compared and analyzed,the albendazole resistance related SNP genes were screened,and their structures were predicted and analyzed by bioinformatics.PCR and RT-PCR amplification and sequence analysis of the selected molecular marker genes of albendazole resistance were carried out,and the detection method of albendazole resistance of Haemonchus contortus was studied.The results are as follows:(1)The transcriptome SNP data of albendazole sensitive and drug-resistant strains of Haemonchus contortus were analyzed,a total of 1154 SNP genes with the same mutation were screened from 5 drug-resistant strains.After GO and KEGG enrichment,770 SNP genes were enriched to 30 GO terms.After KEGG enrichment of 1154 genes,there were20 significant enrichment pathways.(2)Compared with sensitive strains,albendazole resistant molecular marker genes were screened based on the same mutation of five drug-resistant strains,and their protein structure,physical and chemical properties,protein secondary and tertiary structure were analyzed.Results: HCON-00021530(2865T-A),HCON-00109050(360T-A),HCON-00024630(1032C-G)had significant changes in molecular formula,amino acid content,secondary structure and tertiary structure of protein,which can be used as drug resistance molecular marker genes.(3)The genomic DNA and total RNA of eggs of 20 clinical drug-resistant strains were used as templates,PCR and RT-PCR were used to amplified and sequenced the albendazole resistance related SNP genes HCON-00021530,HCON-00109050,HCON-00024630,HCON-00037800 and Isotype-Ⅰ β-tubulin genes of Haemonchus contortus,and RT-PCR and PCR molecular biology detection methods of insect eggs were established.