Drug susceptibility testing methods of antimalarial agents

Maji, Ardhendu Kumar

doi:10.4103/2229-5070.248695

Cited by 14 publications

(7 citation statements)

References 34 publications

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“… 41 Studying in vitro is more rapid and accurate for observing parasite’s fragility due to antimalarial agents, without any disruption from the host factor. 42 …”

Section: Discussionmentioning

confidence: 99%

Fractions 14 and 36K of Metabolite Extract Streptomyces hygroscopicus subsp. Hygroscopicus Have Antimalarial Activities Against Plasmodium berghei in vitro

Fitri¹,

Endharti²,

Abidah³

et al. 2023

IDR

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Purpose The study was conducted to investigate the effectivity and the cytotoxicity of fractions 14 and 36K of metabolite extract of Streptomyces hygroscopicus subsp. Hygroscopicus as an antimalarial compounds against Plasmodium berghei in vitro. Methods Fractions 14 and 36K of metabolite extract of Streptomyces hygroscopicus subsp. Hygroscopicus produced by the fractionation process utilizing the Flash Column Chromatography (FCC) BUCHI Reveleris ® PREP. Plasmodium berghei culture was used to assess the antimalarial activity of fractions 14 and 36K. Parasite densities and the ability of parasite growth were determined under microscopic. The cytotoxicity of the fractions was assessed using MTT assays on the MCF-7 cell line. Results Streptomyces hygroscopicus subsp. Hygroscopicus fractions 14 and 36K have antimalarial activity against Plasmodium berghei , with fraction 14 having the more potent activity. The percentage of Plasmodium berghei -infected erythrocytes was decreased as well as the increase of fraction concentration. Fraction 14 has the highest inhibition of parasite growth at a concentration of 156,25 μg/mL, with an inhibition percentage of 67.73% (R 2 = 0.953, p = 0.000). IC 50 of fractions 14 and 36K were found at 10.63 μg/mL and 135,91 μg/mL, respectively. The fractions caused morphological damage in almost all asexual stages of the parasite. Both fractions were not toxic against MCF-7, indicating that the fractions have a safe active metabolite. Conclusion Fractions 14 and 36K of metabolite extract Streptomyces hygroscopicus subsp. Hygroscopicus contains non-toxic compounds that could damage the morphology and inhibit the growth of Plasmodium berghei in vitro.

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“… 41 Studying in vitro is more rapid and accurate for observing parasite’s fragility due to antimalarial agents, without any disruption from the host factor. 42 …”

Section: Discussionmentioning

confidence: 99%

Fractions 14 and 36K of Metabolite Extract Streptomyces hygroscopicus subsp. Hygroscopicus Have Antimalarial Activities Against Plasmodium berghei in vitro

Fitri¹,

Endharti²,

Abidah³

et al. 2023

IDR

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“…Determining the antimalarial activity of compounds in S. c umini L fruit can be made by in vitro. In vitro antimalarial testing is a test that does not involve the intervention of the host physiological factor, so the results obtained are the result of a parasite that is directly exposed to the test material and can be examined for inhibition of growth and maturation into schizont ( Maji, 2018 ). The result of in vitro testing on malarial material can be determined based on the value of 50 concentration inhibitors (IC 50 ), so this research was conducted with the fraction of S. cumini L fruit as in vitro antimalarial assay against Plasmodium falciparum 3D7 strain to determine the IC 50 value.…”

Section: Introductionmentioning

confidence: 99%

In Vitro antimalarial activity of Syzygium cumini fruit fraction

Maslachah,

Purwitasari

2023

Open Vet J

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Background: Malaria is still one of the most severe public health problems worldwide. The development of treatment, prevention, and control of malaria is one of the substantial problems in the world. Aims: To investigate the in vitro antimalarial activity of Syzygium cumin methanol fruit fraction. Methods: Syzygium cumini L fruit powder was macerated with methanol (PA) and the extract obtained was fractionated using the liquid-liquid partition method with n-hexane, ethyl acetate, butanol, chloroform, methanol, and water solvents. In vitro antimalarial assay was conducted using the culture of P. falciparum 3D7 strain culture that had reached > 5% growth and was examined for IC 50 values using a 24-well microplate in duplicate. Each treatment and control well contained 1080 µl of complete media. Well number 1 was added with 120 µl fraction, then the solution was diluted until it reached 0.01,0.1, 1,10,100 µg/ml the final concentration in the microtiter well. The control only contained complete media and infected erythrocytes without the addition of anti-malarial drugs. The microplate was incubated for 48 hours. After 48 hours a thin blood smear was made fixed with methanol and stained with 20% Giemsa for 20 minutes to determine the IC 50 value by plotting sample concentrations and percentage of parasitaemia in excel. Results: The IC50 values of ethyl acetate fraction, n.hexane fraction, butanol fraction, and water fraction were 1.189 µg/ml, 76.996 µg/ml, 1,769 µg/ml, and 15.058 µg/ml respectively. Whereases the IC50 values of C1 fraction (mix fraction from chloroform: methanol 100:0 and 90:10) and C4 fraction (mix fraction from chloroform: methanol 20:80, 10:90 and 0:100) were 100.126 µg/ml and 1.015 µg/ml respectively. The results showed that the IC50 value of ethyl acetate, butanol, and C4 fraction were lower than 10µg/ml and were considered as good activity (strong antimalarial activity). Conclusion: The ethyl acetate, butanol and C4 subfraction from Syzygium cumini fruit have the potential to be developed as an antimalarial agent.

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“…The methods for testing P. vivax resistance to an antimalarial drug, which include both in vivo and in vitro and molecular marker detection, were always introduced from assessing drug resistance in P. falciparum [30,31]. However, the in vivo accuracy of evaluating the efficacy of anti-malarial drug already faces a challenge due to the difficulty to distinguishing between treatment failure and these infection events, such as recurrence or new infection [32,33].…”

Section: Introductionmentioning

confidence: 99%

Molecular surveillance of chloroquine resistance in Plasmodium vivax isolates from malaria cases in Yunnan Province of China using pvcrt-o gene polymorphisms

Ding,

Dong,

Deng

et al. 2023

Malar J

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Background The efficacy of chloroquine treatment for vivax malaria has been rarely evaluated due to a lack of an appropriate testing method. The objective of this study was to conduct molecular monitoring of chloroquine resistance in Plasmodium vivax strains from vivax malaria patients in Yunnan Province, focusing on the analysis of polymorphism in the P. vivax chloroquine resistance transporter protein orthologous gene (pvcrt-o). Methods In accordance with the principles of a cohort study, blood samples were collected from malaria cases diagnosed with a P. vivax mono-infection in Yunnan Province from 2020 to 2022. Segmental PCR was used to amplify the whole pvcrt-o gene in the blood samples and their products were subsequently sequenced. The sequencing data were arranged to obtain the full coding DNA sequence (CDS) as well as the gene’s promoter region sequences. The CDSs were aligned with the reference sequence (XM_001613407.1) of the P. vivax SalI isolate to identify the mutant loci. Results From a total of 375 blood samples taken from vivax malaria cases, 272 both whole gene CDSs (1272–1275 bp) and promoter DNA sequences (707 bp) of pvcrt-o gene were obtained. Among the whole CDSs, there were 7 single nucleotide polymorphic sites in which c.7 A>G was the minor allele frequency (MAF) site with 4.4% (12/272) detection rate. The mutation detection rate showed a significant decrease from 9.8% (10/102) in 2020 to 1.1% (1/92) in 2021 and 1.3% (1/78) in 2022, indicating statistical significance (χ2 = 11.256, P < 0.05). Among the identified 12 haplotypes, the majority of which were wild type (75.7%; 206/272). These four mutant haplotypes (Hap_3, Hap_5, Hap_9, and Hap_10) were classified as “K10 insertion type” and accounted for 12.1% (33/272). The detection rate of Hap_3 increased from 1.0% (1/102) in 2020 to 13.0% (12/92) in 2021 and 14.1% (11/78) in 2022, indicating statistical significance. A total of 23.8% (65/272) of the samples exhibited 14 bp (bp) deletions in the promoter region, occurring most frequently in the wild type haplotype (Hap_1) samples at a rate of 28.6% (59/206). Conclusions In recent years in Yunnan Province, a notable proportion of vivax malaria patients are infected by P. vivax strains with a “K10 insertion” and partial sequence deletions in the promoter region of the pvcrt-o gene, necessitating vigilance.

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Drug susceptibility testing methods of antimalarial agents

Cited by 14 publications

References 34 publications

Fractions 14 and 36K of Metabolite Extract Streptomyces hygroscopicus subsp. Hygroscopicus Have Antimalarial Activities Against Plasmodium berghei in vitro

Fractions 14 and 36K of Metabolite Extract Streptomyces hygroscopicus subsp. Hygroscopicus Have Antimalarial Activities Against Plasmodium berghei in vitro

In Vitro antimalarial activity of Syzygium cumini fruit fraction

Molecular surveillance of chloroquine resistance in Plasmodium vivax isolates from malaria cases in Yunnan Province of China using pvcrt-o gene polymorphisms

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