Significant geographical differences in prevalence of mutations associated with Plasmodium falciparum and Plasmodium vivax drug resistance in two regions from Papua New Guinea

Abstract

BackgroundDrug resistance remains a major obstacle to malaria treatment and control. It can arise and spread rapidly, and vary substantially even at sub-national level. National malaria programmes require cost-effective and timely ways of characterizing drug-resistance at multiple sites within their countries.MethodsAn improved multiplexed post-PCR ligase detection reaction—fluorescent microsphere assay (LDR-FMA) was used to simultaneously determine the presence of mutations in chloroquine resistance transporter (crt), multidrug resistance 1 (mdr1), dihydrofolate reductase (dhfr) and dihydropteroate synthase (dhps) genes in Plasmodium falciparum (n = 727) and Plasmodium vivax (n = 574) isolates collected in 2006 from cross-sectional community population surveys in two geographically distinct regions (Madang and East Sepik) of Papua New Guinea (PNG) where strong regional differences in in vivo aminoquinoline and antifolate therapeutic efficacy had previously been observed. Data were compared to those of a follow-up survey conducted in 2010.ResultsDespite some very low parasite densities, the assay successfully amplified all P. falciparum and P. vivax loci in 77 and 69 % of samples, respectively. In 2006, prevalences of pfdhfr (59R-108 N) double mutation/wild type pfdhps haplotype, pfcrt SVMNT haplotype (72S-76T double mutation), and 86Y pfmdr1 mutation all exceeded 90 %. For P. vivax, 65 % carried at least two pvdhfr mutations, 97 % the 647P pvdhps mutation and 54 % the 976F pvmdr1 mutation. Prevalence of mutant haplotypes was higher in Madang than East Sepik for pfcrt SVMNT (97.4 vs 83.3 %, p = 0.001), pfdhfr (59R-108 N) (100 vs 90.6 %, p = 0.001), pvdhfr haplotypes (75.8 vs 47.6 %, p = 0.001) and pvmdr1 976F (71.2 vs 26.2 %, p < 0.001). Data from a subsequent Madang survey in 2010 showed that the prevalence of pfdhps mutations increased significantly from <5 % to >30 % (p < 0.001) as did the prevalence of pvdhfr mutant haplotypes (from 75.8 to 97.4 %, p = 0.012).ConclusionsThis LDR-FMA multiplex platform shows feasibility for low-cost, high-throughput, rapid characterization of a broad range of drug-resistance markers in low parasitaemia infections. Significant geographical differences in mutation prevalence correlate with previous genotyping surveys and in vivo trials and may reflect variable drug pressure and differences in health-care access in these two PNG populations.Electronic supplementary materialThe online version of this article (doi:10.1186/s12936-015-0879-9) contains supplementary material, which is available to authorized users.

Molecular surveillance of Pfcrt and k13 propeller polymorphisms of imported Plasmodium falciparum cases to Zhejiang Province, China between 2016 and 2018

“…Our study confirmed that CVIET was the most common mutation type in infections imported from Africa, similar to previous studies [26,28]. The high frequency of SVMNT in Papua New Guinea was also consistent with other studies [29]. It was demonstrated that the spatial distribution of mutant alleles was mainly related to local drug policy.…”

Molecular Detection of Antimalarial Drug Resistance in Plasmodium vivax from Returned Travellers to NSW, Australia during 2008–2018

“…The present study found the L/I57:R58:M61:T117 mutations in a large number of isolates from Oceania (21/37), which was followed by the Southeast Asian (4/10) and South Asian (2/73) regions. Our findings were consistent with other studies from these regions [5,7,32,[50][51][52]. An increase in treatment failure by 12% occurred in P. vivax cases in PNG when the malaria treatment policy in 2000 changed to recommending the use of SP with either chloroquine or amodiaquine as the first-line drug for uncomplicated malaria [53].…”

“…However, nine isolates from South Asia (6/73), Southeast Asia (2/10) and Oceania (1/37) had the G383:G55 double mutations, which are linked to sulfadoxine resistance in P. vivax [5,7,58]. The current results correlate with the findings made in previous studies from these regions [7,31,32,51,52,55,59]. Some studies [47,60] showed that the mutations in the Pvdhps gene were more than twice as likely to emerge in isolates with multiple mutations in the Pvdhfr gene.…”

“…The finding of the F976:L1076 mutations in Oceania is, however, different from earlier studies in PNG between 2006 and 2011; these studies showed the presence of the F976 mutation only [30][31][32]. This suggests that there was a rise in the number of chloroquine resistance markers since the first reported case of resistant P. vivax in the region, even though chloroquine has not been recommended in the antimalarial drug policy since 2009 [33].…”

Markers of sulfadoxine–pyrimethamine resistance in Eastern Democratic Republic of Congo; implications for malaria chemoprevention

“…For all qPCR positive samples, nested PCR amplification was done for dhps; for a sub-set of samples where dhps amplification was successful, dhfr was also amplified (n = 387). Primers were based on previous publications [16,17] and are presented in Table 1. PCR conditions were 94 °C × 3 min, followed by 40 rounds of 94 °C × 1 min → 51 °C × 2 min → 72 °C × 1 min, and a final round of 72 °C for 10 min.…”