Two novel mutations of pfdhps K540T and I588F, affecting sulphadoxine-pyrimethamine-resistant response in uncomplicated falciparum malaria at Banjar district, South Kalimantan Province, Indonesia.

Sukmawati Basuki,Yoes P Dachlan,Budiono Budiono,Haruki Uemura,Sugeng Riyanto,Fitriah Fitriah

doi:10.1186/1475-2875-13-135

Abstract

BackgroundMutations in pfdhfr and pfdhps genes have been shown to associate with sulphadoxine-pyrimethamine (SP) resistance of Plasmodium falciparum parasites. However, pfdhfr, pfdhps genotypes and the correlations to SP treatment outcome in Indonesia has not yet been well analysed.MethodsAfter obtaining informed consent, 61 uncomplicated falciparum malaria patients were recruited in Banjar district, South Kalimantan Province, Indonesia, from October 2009 to August 2010. They were treated by a single oral dose of SP and its effects on clinical and parasitological status were followed until day 28 after treatment. Occasionally, a thick smear blood film for microscopy observation and blood spot on a filter paper for pfdhfr and pfdhps genotype analysis were collected.ResultsPfdhfr and pfdhps genotypes from 24 P. falciparum-infected patients consisting of adequate clinical parasitological response (ACPR) (n = 6; 25.0%) and early treatment failure (ETF) (n = 10; 41.7%) or late parasitological failure (LPF) (n = 8; 33.3%) were obtained by sequencing. Two novel mutations of pfdhps gene, K540T and I588F, were determined in ten and five isolates, respectively. These mutations were present in the pfdhfr/pfdhps combined haplotypes of ANRNI/SGTGA (n = 6), ANRNL/SGTGA (n = 4), and ANRNI/SGEAA(588F) (n = 5), (mutation codons are bold typed); these haplotypes were mostly belonging to parasitological failure (ETF or LPF). The parasites acquiring five mutations in pfdhfr/pfdhps haplotypes and four mutations with additional I588F did not respond adequately to SP treatment.ConclusionMany of Plasmodium falciparum infected patients in Banjar district, South Kalimantan, Indonesia did not respond adequately to SP treatment and these low ineffectiveness of SP in this area was associated with two novel mutations of pfdhps, K540T and I588F.

Highlights

Mutations in pfdhfr and pfdhps genes have been shown to associate with sulphadoxine-pyrimethamine (SP) resistance of Plasmodium falciparum parasites
High rates of chloroquine resistance in falciparum and vivax malaria have been reported in Indonesia and sulphadoxine-pyrimethamine (SP) resistance in falciparum malaria was discovered in several endemic areas from 1981 to 1995
The detected parasites were few in number in many cases and it makes such cases difficult to determine whether early treatment failure (ETF) or late parasitological failure (LPF) clearly, some correlations between pfdhfr/pfdhps haplotypes and SP sensitivity were observed

Summary

Introduction

Mutations in pfdhfr and pfdhps genes have been shown to associate with sulphadoxine-pyrimethamine (SP) resistance of Plasmodium falciparum parasites. In vitro and in vivo SP resistance were found in four provinces: Central Java, East Timor, South Sulawesi, and Papua in 1991 by Tjitra et al [3] and in vitro resistance was confirmed by using field isolates obtained from 11 malaria-endemic provinces, including East Kalimantan Province [3,4]. In spite of these reports, people in several endemic areas still commonly use SP when they are suffering from malaria because of its effectiveness, fewer side effects, low cost, and the single oral dose treatment. SP is provided for chemoprophylaxis [5]

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