Assessment of Plasmodium falciparum anti-malarial drug resistance markers in pfcrt and pfmdr1 genes in isolates from Honduras and Nicaragua, 2018–2021

Fontecha, Gustavo; Pinto, Alejandra; Archaga, Osman; Betancourth, Sergio; Escober, Lenin; Henríquez, Jessica; Valdivia, Hugo O.; Montoya, Alberto; Mejía, Rosa Elena

doi:10.1186/s12936-021-03977-8

Cited by 7 publications

(5 citation statements)

References 59 publications

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“…Pfmdr1 gene was amplified to identify mutation at codon N86Y according to the protocol described by Fontecha et al [ 24 ] with certain modifications. Briefly, both primary and nested PCRs were conducted in a total reaction volume of 18.0 µl consisting of 11.4 µl ddH 2 O, 1.0 µl of each forward and reverse primers (Additional file 1 ), 3.6 µl 5X FIREPol Master Mix and 1.0 µl DNA template.…”

Section: Molecular Analysismentioning

confidence: 99%

High prevalence of Pfcrt 76T and Pfmdr1 N86 genotypes in malaria infected patients attending health facilities in East Shewa zone, Oromia Regional State, Ethiopia

Hassen

Alemayehu

Dinka

et al. 2022

Malar J

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Background Plasmodium falciparum resistance to series of anti-malarial drugs is a major challenge in efforts to control and/or eliminate malaria globally. In 1998, following the widespread of chloroquine (CQ) resistant P. falciparum, Ethiopia switched from CQ to sulfadoxine–pyrimethamine (SP) and subsequently in 2004 from SP to artemether–lumefantrine (AL) for the treatment of uncomplicated falciparum malaria. Data on the prevalence of CQ resistance markers after more than two decades of its removal is important to map the selection pressure behind the targets codons of interest. The present study was conducted to determine the prevalence of mutations in Pfcrt K76T and Pfmdr1 N86Y codons among malaria-infected patients from Adama, Olenchiti and Metehara sites of East Shewa zone, Oromia Regional State, Ethiopia. Methods Finger-prick whole blood samples were collected on 3MM Whatman ® filter papers from a total of 121 microscopically confirmed P. falciparum infected patients. Extraction of parasite DNA was done by Chelex-100 method from dried blood spot (DBS). Genomic DNA template was used to amplify Pfcrt K76T and Pfmdr1 N86Y codons by nested PCR. Nested PCR products were subjected to Artherobacter protophormiae-I (APoI) restriction enzyme digestion to determine mutations at codons 76 and 86 of Pfcrt and Pfmdr1 genes, respectively. Results Of 83 P. falciparum isolates successfully genotyped for Pfcrt K76T, 91.6% carried the mutant genotypes (76T). The prevalence of Pfcrt 76T was 95.7%, 92.5% and 84.5% in Adama, Metehara and Olenchiti, respectively. The prevalence of Pfcrt 76T mutations in three of the study sites showed no statistical significance difference (χ2 = 1.895; P = 0.388). On the other hand, of the 80 P. falciparum samples successfully amplified for Pfmdr1, all carried the wild-type genotypes (Pfmdr1 N86). Conclusion Although CQ officially has been ceased for the treatment of falciparum malaria for more than two decades in Ethiopia, greater proportions of P. falciparum clinical isolates circulating in the study areas carry the mutant 76T genotypes indicating the presence of indirect CQ pressure in the country. However, the return of Pfmdr1 N86 wild-type allele may be favoured by the use of AL for the treatment of uncomplicated falciparum malaria.

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Section: Molecular Analysismentioning

confidence: 99%

High prevalence of Pfcrt 76T and Pfmdr1 N86 genotypes in malaria infected patients attending health facilities in East Shewa zone, Oromia Regional State, Ethiopia

Hassen

Alemayehu

Dinka

et al. 2022

Malar J

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“…The approach proposed in this study has the limitation that genotype D would not guarantee with absolute certainty that it is not a case imported from South America, Africa, or even Papua New Guinea, and it would be necessary to apply other molecular markers to resolve the situation. A marker that could be used for this purpose is pfcrt , since strains of P. falciparum from Honduras, Guatemala, and Nicaragua show a wild-type genotype sensitive to chloroquine, very rarely found in other regions [ 6 ]. One of the main limitations of this study is the low number of local samples analyzed, but taking into account the drastic decrease in malaria cases in Honduras and the low genetic diversity demonstrated in parasite populations [ 41 ], the probability that most or all of the circulating strains are of genotype D is high.…”

Section: Discussionmentioning

confidence: 99%

“…A 94 bp segment encompassing the polymorphic region of domain 2 of the pfs47 gene was amplified. DNA was extracted from 32 blood samples deposited on filter paper and previously diagnosed as cases of falciparum malaria from Honduras and Nicaragua [ 6 ].…”

Section: Methodsmentioning

confidence: 99%

“…strains from South America or the Old World represents an additional challenge for the public health systems of countries with limited resources such as Honduras. The introduction of virulent P. falciparum strains that require a treatment scheme different from that indicated by the National Standard with chloroquine and primaquine [ 6 ] is a risk for the management of imported cases.…”

Section: Introductionmentioning

confidence: 99%

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A PCR-RFLP Technique to Assess the Geographic Origin of Plasmodium falciparum Strains in Central America

et al. 2022

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The elimination of malaria requires strengthening diagnosis and offering adequate and timely treatment. Imported cases of falciparum malaria represent a major challenge for pre-elimination areas, such as Central America, where chloroquine and primaquine continue to be used as first-line treatment. The pfs47 gene has been previously described as a precise molecular marker to track the geographic origin of the parasite. The aim of this study was to design a simple and low-cost technique using the polymorphic region of pfs47 to assess the geographic origin of P. falciparum strains. A PCR-RFLP technique was developed and evaluated using the MseI enzyme that proved capable of discriminating, with reasonable precision, the geographical origin of the parasites. This method could be used by national surveillance laboratories and malaria elimination programs in countries such as Honduras and Nicaragua in cases of malaria where an origin outside the Central American isthmus is suspected.

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“…2 Despite several reports of imported chloroquine-resistant P. falciparum infections in the area and the detection of PfCRT mutations in 0.8–1.3% of samples examined, such an increment is not related to the introduction of drug-resistant malaria but to health threats related to social, political, economic and gender inequalities. 10 , 11 , 12…”

Section: Malaria In the Americas– A Snapshotmentioning

confidence: 99%

Migrants in transit across Central America and the potential spread of chloroquine resistant malaria–a call for action

Higuita

Franco‐Paredes

Henao‐Martínez

et al. 2023

The Lancet Regional Health - Americas

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Assessment of Plasmodium falciparum anti-malarial drug resistance markers in pfcrt and pfmdr1 genes in isolates from Honduras and Nicaragua, 2018–2021

Cited by 7 publications

References 59 publications

High prevalence of Pfcrt 76T and Pfmdr1 N86 genotypes in malaria infected patients attending health facilities in East Shewa zone, Oromia Regional State, Ethiopia

High prevalence of Pfcrt 76T and Pfmdr1 N86 genotypes in malaria infected patients attending health facilities in East Shewa zone, Oromia Regional State, Ethiopia

A PCR-RFLP Technique to Assess the Geographic Origin of Plasmodium falciparum Strains in Central America

Migrants in transit across Central America and the potential spread of chloroquine resistant malaria–a call for action

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