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      Surveillance of molecular markers of antimalarial drug resistance in Plasmodium falciparum and Plasmodium vivax in Federally Administered Tribal Area (FATA), Pakistan

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          Abstract

          ABSTRACT This molecular epidemiological study was designed to determine the antimalarial drug resistance pattern, and the genetic diversity of malaria isolates collected from a war-altered Federally Administered Tribal Area (FATA), in Pakistan. Clinical isolates were collected from Bajaur, Mohmand, Khyber, Orakzai and Kurram agencies of FATA region between May 2017 and May 2018, and they underwent DNA extraction and amplification. The investigation of gene polymorphisms in drug resistance genes (dhfr, dhps, crt, and mdr1) of Plasmodium falciparum and Plasmodium vivax was carried out by pyrosequencing and Sanger sequencing, respectively. Out of 679 PCR-confirmed malaria samples, 523 (77%) were P. vivax, 121 (18%) P. falciparum, and 35 (5%) had mixed-species infections. All P. falciparum isolates had pfdhfr double mutants (C59R+S108N), while pfdhfr/pfdhps triple mutants (C59R+S108N+A437G) were detected in 11.5% of the samples. About 97.4% of P. falciparum isolates contained pfcrt K76T mutation, while pfmdr1 N86Y and Y184F mutations were present in 18.2% and 10.2% of the samples. P. vivax pvdhfr S58R mutation was present in 24.9% of isolates and the S117N mutation in 36.2%, while no mutation in the pvdhps gene was found. Pvmdr1 F1076L mutation was found in nearly all samples, as it was observed in 98.9% of isolates. No significant anti-folate and chloroquine resistance was observed in P. vivax; however, mutations associated with antifolate-resistance were found, and the chloroquine-resistant gene has been observed in 100% of P. falciparum isolates. Chloroquine and sulphadoxine-pyrimethamine resistance were found to be high in P. falciparum and low in P. vivax. Chloroquine could still be used for P. vivax infection but need to be tested in vivo, whereas a replacement of the artemisinin combination therapy for P. falciparum appears to be justified.

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          Spread of artemisinin resistance in Plasmodium falciparum malaria.

          Elizabeth A Ashley, Mehul Dhorda, Rick Fairhurst … (2014)
          Artemisinin resistance in Plasmodium falciparum has emerged in Southeast Asia and now poses a threat to the control and elimination of malaria. Mapping the geographic extent of resistance is essential for planning containment and elimination strategies. Between May 2011 and April 2013, we enrolled 1241 adults and children with acute, uncomplicated falciparum malaria in an open-label trial at 15 sites in 10 countries (7 in Asia and 3 in Africa). Patients received artesunate, administered orally at a daily dose of either 2 mg per kilogram of body weight per day or 4 mg per kilogram, for 3 days, followed by a standard 3-day course of artemisinin-based combination therapy. Parasite counts in peripheral-blood samples were measured every 6 hours, and the parasite clearance half-lives were determined. The median parasite clearance half-lives ranged from 1.9 hours in the Democratic Republic of Congo to 7.0 hours at the Thailand-Cambodia border. Slowly clearing infections (parasite clearance half-life >5 hours), strongly associated with single point mutations in the "propeller" region of the P. falciparum kelch protein gene on chromosome 13 (kelch13), were detected throughout mainland Southeast Asia from southern Vietnam to central Myanmar. The incidence of pretreatment and post-treatment gametocytemia was higher among patients with slow parasite clearance, suggesting greater potential for transmission. In western Cambodia, where artemisinin-based combination therapies are failing, the 6-day course of antimalarial therapy was associated with a cure rate of 97.7% (95% confidence interval, 90.9 to 99.4) at 42 days. Artemisinin resistance to P. falciparum, which is now prevalent across mainland Southeast Asia, is associated with mutations in kelch13. Prolonged courses of artemisinin-based combination therapies are currently efficacious in areas where standard 3-day treatments are failing. (Funded by the U.K. Department of International Development and others; ClinicalTrials.gov number, NCT01350856.).
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            Mutations in the P. falciparum digestive vacuole transmembrane protein PfCRT and evidence for their role in chloroquine resistance.

            L M Ursos, X Su, B Naudé … (2000)
            The determinant of verapamil-reversible chloroquine resistance (CQR) in a Plasmodium falciparum genetic cross maps to a 36 kb segment of chromosome 7. This segment harbors a 13-exon gene, pfcrt, having point mutations that associate completely with CQR in parasite lines from Asia, Africa, and South America. These data, transfection results, and selection of a CQR line harboring a novel K761 mutation point to a central role for the PfCRT protein in CQR. This transmembrane protein localizes to the parasite digestive vacuole (DV), the site of CQ action, where increased compartment acidification associates with PfCRT point mutations. Mutations in PfCRT may result in altered chloroquine flux or reduced drug binding to hematin through an effect on DV pH.
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              Identification of the four human malaria parasite species in field samples by the polymerase chain reaction and detection of a high prevalence of mixed infections.

              G Snounou, S Viriyakosol, W Jarra … (1993)
              Genus- and species-specific sequences are present within the small subunit ribosomal RNA genes of the four human malaria parasites. Oligonucleotide primer pairs specific to each species were designed for specific amplification by the Polymerase Chain Reaction (PCR), to detect each malaria species. DNA equivalent to 5 microliters of blood was sufficient for the detection of each of the species. Blood samples obtained from 196 patients attending a malaria clinic in Trad province (Thailand) were analyzed. Detection and identification of the parasites, solely by electrophoretic analysis of the PCR products, has proven to be more sensitive and accurate than by routine diagnostic microscopy. A high proportion of mixed species infections were brought to light by the PCR assay. Implications for medical treatment and epidemiological studies are discussed.
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                Author and article information

                Journal
                Journal ID (publisher-id): rimtsp
                Title: Revista do Instituto de Medicina Tropical de São Paulo
                Abbreviated Title: Rev. Inst. Med. trop. S. Paulo
                Publisher: Instituto de Medicina Tropical de São Paulo (São Paulo, SP, Brazil )
                ISSN (Electronic): 1678-9946
                Publication date (Print and electronic): 2021
                Volume: 63
                Electronic Location Identifier: e59
                Affiliations
                [2] Haripur Khyber Pakhtunkhwa orgnameThe University of Haripur orgdiv1Department of Medical Laboratory Technology Pakistan
                [3] Quetta Balochistan orgnameSandeman Provincial Hospital orgdiv1Department of Surgery Pakistan
                [1] Gujrat Punjab orgnameUniversity of Gujrat orgdiv1Department of Biochemistry & Biotechnology Pakistan
                [4] Rawalpindi Pujab orgnameNational University of Medical Sciences orgdiv1Department of Biological Sciences Pakistan
                [5] Haripur Khyber Pakhtunkhwa orgnameThe University of Haripur orgdiv1Department of Microbiology Pakistan
                Article
                Publisher ID: S0036-46652021000100231 Publisher ID: S0036-4665(21)06300000231
                DOI: 10.1590/s1678-994620216305
                SO-VID: 1f92ae50-490a-4457-b23c-ce0b69dcba6b
                License:

                This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.

                History
                Date received : 30 April 2021
                Date accepted : 16 June 2021
                Page count
                Figures: 0, Tables: 0, Equations: 0, References: 40, Pages: 0
                Product

                SciELO Brazil

                Categories
                Subject: Original Article

                Keywords: Antimalarial drug resistance,Malaria,FATA,Plasmodium falciparum,Plasmodium vivax,Pakistan
                Data availability:
                Keywords: Antimalarial drug resistance, Malaria, FATA, Plasmodium falciparum, Plasmodium vivax, Pakistan

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