Development of a Tetra ARMS-PCR based method for genotyping the A296S position in An. stephensi to monitor the resistance to GABA Receptor based insecticides
عنوان دوره: اولین کنگره بین المللی حشره شناسی ایران
نویسندگان
چکیده
Malaria is one of the most important infectious diseases throughout the world. The main carrier of the malaria in Iran is Anopheles Stephensi. One of the strategies for vector controlling is insecticide application. A type of insecticides acts on GABA receptor and nervous system. Therefore, the aim of this study is design and development of a rapid, sensitive and accurate technique based on Tetra ARMS-PCR method for diagnosis the mutation which is related to resistance against effective toxins on GABA receptor in An. stephensi.
In this study, 100 mosquitoes were sampled by total catch method from Chabahar of Sistan and Balochestan province. DNA was extracted by using of MBST DNA extract kit. Then, Tetra ARMS-PCR was performed for A296S position. Finally, the accuracy of the results of Tetra ARMS-PCR method were evaluated by sequencing the samples by different genotypes.
In this study, the A296S position was genotyped in 100 samples and it was revealed that the genotype of 80 samples is heterozygote and 20 samples is mutant. No wild allele was observed for A296S in collected samples. The comparison of our results with sequencing revealed that the accuracy of our developed method is 100%.
Therefore, our results showed that the our developed method is reliable, sensitive and accurate and it can be suggested as an alternative method instead of RFLP for molecular monitoring of A296S position.
In this study, 100 mosquitoes were sampled by total catch method from Chabahar of Sistan and Balochestan province. DNA was extracted by using of MBST DNA extract kit. Then, Tetra ARMS-PCR was performed for A296S position. Finally, the accuracy of the results of Tetra ARMS-PCR method were evaluated by sequencing the samples by different genotypes.
In this study, the A296S position was genotyped in 100 samples and it was revealed that the genotype of 80 samples is heterozygote and 20 samples is mutant. No wild allele was observed for A296S in collected samples. The comparison of our results with sequencing revealed that the accuracy of our developed method is 100%.
Therefore, our results showed that the our developed method is reliable, sensitive and accurate and it can be suggested as an alternative method instead of RFLP for molecular monitoring of A296S position.
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