Abstract
BACKGROUND AND OBJECTIVE Brazil is responsible for a large number of Plasmodium vivax cases in America. Given the emergence of P. vivax parasites resistant to chloroquine and the effectiveness of antifolates in vivax malaria treatment together with a correlation between mutations in P. vivax dhfr and dhps genes and SP treatment failure, the point mutations in these genes were investigated.METHODS Blood samples from 54 patients experiencing vivax malaria symptomatic episodes in the Amazonian Region were investigated. Genomic DNA was extracted using a DNA extraction kit (QIAGENTM). Nested polymerase chain reaction (PCR) amplification was carried out followed by Sanger sequencing to detect single nucleotide polymorphisms (SNPs).FINDINGS All tested isolates showed non-synonymous mutations in pvdhfr gene: 117N (54/54, 100%) and 58R (25/54, 46%). Double mutant allele 58R/117N (FRTNI, 28%) was the most frequent followed by triple mutant alleles (58R/117N/173L, FRTNL, 11%; 58R/61M/117N, FRMNI, 5% 117N/173L, FSTNL, 4%) and quadruple mutant allele (58R/61M/117N/173L, FRMNL, 2%). A single mutation was observed at codon C383G in pvdhps gene (SGKAV, 48%).CONCLUSION No evidence of molecular signatures associated with P. vivax resistance to SP was observed in the Brazilian samples.
Highlights
MethodsBlood samples from 54 patients experiencing vivax malaria symptomatic episodes in the Amazonian Region were investigated
AND OBJECTIVE Brazil is responsible for a large number of Plasmodium vivax cases in America
No evidence of molecular signatures associated with P. vivax resistance to SP was observed in the Brazilian samples
Summary
Blood samples from 54 patients experiencing vivax malaria symptomatic episodes in the Amazonian Region were investigated. Parasites isolates and DNA extraction - Blood samples from Amazon Region (Acre, Amapá, Amazonas, Rondônia and Pará) were collected from 54 patients presenting with vivax malaria from 2010 to 2016 at the Laboratório de Doenças Febris Agudas, INI-IPEC, Fiocruz, the Reference Clinical Laboratory for Malaria in the ExtraAmazon to the Brazilian Ministry of Health. All the clinical isolates were diagnosed as single P. vivax infections by light microscopic examination of Giemsa’s solutionstained blood smears and by P. vivax cysteine-proteinase target gene polymerase chain reaction (PCR).(10) The parasitaemia ranged from 960 to 19160 parasites/μL. Patients were treated with CQ plus PQ, according to the Brazilian Ministry of Health recommendation for uncomplicated vivax malaria treatment and were followed up to 42 days.
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