Multiple PCR primer for expanding plasmodium falciparum drug resistance-related gene and method

Abstract

The invention relates to a plasmodium falciparum drug resistance related gene multiple PCR amplification method, wherein, plasmodium falciparum genome DNA is adopted as template; a multiple PCR primer set designed by the invention is adopted for monotube PCR reaction to realize amplification of five specific gene (plasmodium falciparum chloroquine resistance transport protein gene Pfcrt, plasmodium falciparum multidrug-resistance gene Pfmdrl, plasmodium falciparum dihydropteroic synthetase gene Pfdhps, plasmodium falciparum dihyrofolate reductase gene Pfdhfr and plasmodium falciparum adenosine triphosphatase Sixth subunit gene PfATPase6) target sequences; moreover, the amplification product covers the currently known 21 plasmodium falciparum drug-resistance related SNP sites. With fewer operational procedures, the invention is quick and convenient and saves cost; moreover, the invention which is particularly suitable for high-throughput genotype analysis is sure to accelerate the wide application of plasmodium falciparum resistance related SNP in field monitoring.

Description

technical field [0001] The invention relates to an amplification technology of a gene related to drug resistance of Plasmodium falciparum, and belongs to the technical field of nucleotide determination or detection methods. Background technique [0002] The prevalence of malaria is severe worldwide, especially the harm of falciparum malaria. According to the report of the World Health Organization, about 1 million people die of falciparum malaria every year in the world, not including indirect deaths. Falciparum malaria is prevalent in Yunnan and Hainan provinces in my country, and deaths are reported every year. After the 1950s, Plasmodium falciparum began to develop resistance to antimalarial drugs. In recent years, the resistance situation has become increasingly severe, from low resistance to high resistance, from single resistance to multiple resistance, and the speed of resistance development Gradually speed up. Accurate and rapid monitoring of the distribution and p...

AI Technical Summary

Problems solved by technology

[0149] Aiming at the disadvantages of cumbersome operation, time-consuming and labor-intensive traditional amplification methods of Plasmodium falciparum drug resistance-related genes, the present invention provides an efficient and convenient multiplex PCR amplification primer and method for Plasmodium falciparum drug resistance-related genes, realizing one-time The reaction completed the amplification of 5 main resistance-related genes (respectively reflecting the resistance of Plasmodium falciparum to chloroquine, mefloquine, pyrimethamine, sulfonamides and artemisinin and other drugs), and the amplification products covered 21 Resistance-related SNPs provide important technical support for subsequent SNP analysis, such as DNA sequencing, RFLP, PCR-ELISA and SNP chip detection methods, thereby significantly improving the detection efficiency of Plasmodium falciparum drug resistance-related SNPs. The extensive development of resistance molecular marker monitoring lays a technical foundation

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