Selection and Spread of Artemisinin-Resistant Alleles in Thailand Prior to the Global Artemisinin Resistance Containment Campaign

Abstract

The recent emergence of artemisinin resistance in the Greater Mekong Subregion poses a major threat to the global effort to control malaria. Tracking the spread and evolution of artemisinin-resistant parasites is critical in aiding efforts to contain the spread of resistance. A total of 417 patient samples from the year 2007, collected during malaria surveillance studies across ten provinces in Thailand, were genotyped for the candidate Plasmodium falciparum molecular marker of artemisinin resistance K13. Parasite genotypes were examined for K13 propeller mutations associated with artemisinin resistance, signatures of positive selection, and for evidence of whether artemisinin-resistant alleles arose independently across Thailand. A total of seven K13 mutant alleles were found (N458Y, R539T, E556D, P574L, R575K, C580Y, S621F). Notably, the R575K and S621F mutations have previously not been reported in Thailand. The most prevalent artemisinin resistance-associated K13 mutation, C580Y, carried two distinct haplotype profiles that were separated based on geography, along the Thai-Cambodia and Thai-Myanmar borders. It appears these two haplotypes may have independent evolutionary origins. In summary, parasites with K13 propeller mutations associated with artemisinin resistance were widely present along the Thai-Cambodia and Thai-Myanmar borders prior to the implementation of the artemisinin resistance containment project in the region.

Author Summary

The Plasmodium falciparum parasites that cause malaria are evolving resistance to our most effective and potent anti-malarial drugs available, called artemisinins. Currently, artemisinin resistance is emerging in a number of countries in the Greater Mekong Subregion, including Cambodia, Thailand, Myanmar, and Vietnam. Historically, the Thai-Cambodia border region has been an epicenter of resistance to several anti-malarial drugs. To prevent the spread of artemisinin resistant parasites from the Greater Mekong Subregion, a global artemisinin resistance project was initiated in 2009. Here, we show that artemisinin resistance associated mutation in the K13 gene were widely present throughout Thailand, as early as 2007, primarily along the Thai-Cambodia and Thai-Myanmar border regions. Additional data based on microsatellite markers suggests that the most commonly found K13 C580Y allele may have two recent independent origins in Thailand, on the borders of Cambodia and Myanmar.