Quantification of Plasmodium falciparum HRP-2 as an alternative method to [3H]hypoxanthine incorporation to measure the parasite reduction ratio in vitro
Copyright © 2023 Elsevier Ltd and International Society of Antimicrobial Chemotherapy. All rights reserved..
In the absence of a highly efficacious vaccine, chemotherapy remains the cornerstone to control malaria morbidity and mortality. The threat of the emergence of parasites resistant to artemisinin-based combination therapies highlights the need for new antimalarial drugs ideally with superior properties. The killing rate reflects the speed of action of antimalarial drugs, which can be measured in vitro through the parasite reduction ratio (PRR) assay to shortlist interesting candidates. As a standard, the in vitro PRR assay is performed by measuring [3H]hypoxanthine incorporation of Plasmodium falciparum. This methodology is restricted to specialised laboratories owing to the handling of radioactive material. In this work, we describe a sandwich enzyme-linked immunosorbent assay to detect P. falciparum histidine-rich protein 2 (HRP-2) as an alternative methodology to assess the PRR. We first validated the methodology with established antimalarial drugs (artesunate, chloroquine, pyrimethamine and atovaquone) by comparing our results with previous results of the [3H]hypoxanthine incorporation readout provided by an expert laboratory, and subsequently assessed the speed of action of four new antimalarial candidates (compound 22, chlorotonil A, boromycin and ivermectin). The HRP-2 PRR assay achieved comparable results to the [3H]hypoxanthine incorporation readout in terms of parasite growth rate over time, lag phase and parasite clearance time. In addition, parasite growth following drug exposure was quantified after 7, 14, 21 and 28 days of recovery time. In conclusion, the PRR assay based on HRP-2 is similar to [3H]hypoxanthine in determining a drug's parasite killing rate and can be widely used in all research laboratories.
| Medienart: |
E-Artikel |
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| Erscheinungsjahr: |
2023 |
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| Erschienen: |
2023 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:62 |
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| Enthalten in: |
International journal of antimicrobial agents - 62(2023), 3 vom: 20. Sept., Seite 106894 |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
de Carvalho, Laís Pessanha [VerfasserIn] |
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| Links: |
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| Themen: |
[(3)H]hypoxanthine |
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| Anmerkungen: |
Date Completed 29.08.2023 Date Revised 29.08.2023 published: Print-Electronic Citation Status MEDLINE |
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| doi: |
10.1016/j.ijantimicag.2023.106894 |
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| funding: |
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| Förderinstitution / Projekttitel: |
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| PPN (Katalog-ID): |
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| 245 | 1 | 0 | |a Quantification of Plasmodium falciparum HRP-2 as an alternative method to [3H]hypoxanthine incorporation to measure the parasite reduction ratio in vitro |
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| 520 | |a Copyright © 2023 Elsevier Ltd and International Society of Antimicrobial Chemotherapy. All rights reserved. | ||
| 520 | |a In the absence of a highly efficacious vaccine, chemotherapy remains the cornerstone to control malaria morbidity and mortality. The threat of the emergence of parasites resistant to artemisinin-based combination therapies highlights the need for new antimalarial drugs ideally with superior properties. The killing rate reflects the speed of action of antimalarial drugs, which can be measured in vitro through the parasite reduction ratio (PRR) assay to shortlist interesting candidates. As a standard, the in vitro PRR assay is performed by measuring [3H]hypoxanthine incorporation of Plasmodium falciparum. This methodology is restricted to specialised laboratories owing to the handling of radioactive material. In this work, we describe a sandwich enzyme-linked immunosorbent assay to detect P. falciparum histidine-rich protein 2 (HRP-2) as an alternative methodology to assess the PRR. We first validated the methodology with established antimalarial drugs (artesunate, chloroquine, pyrimethamine and atovaquone) by comparing our results with previous results of the [3H]hypoxanthine incorporation readout provided by an expert laboratory, and subsequently assessed the speed of action of four new antimalarial candidates (compound 22, chlorotonil A, boromycin and ivermectin). The HRP-2 PRR assay achieved comparable results to the [3H]hypoxanthine incorporation readout in terms of parasite growth rate over time, lag phase and parasite clearance time. In addition, parasite growth following drug exposure was quantified after 7, 14, 21 and 28 days of recovery time. In conclusion, the PRR assay based on HRP-2 is similar to [3H]hypoxanthine in determining a drug's parasite killing rate and can be widely used in all research laboratories | ||
| 650 | 4 | |a Journal Article | |
| 650 | 4 | |a HRP-2 | |
| 650 | 4 | |a Killing rate | |
| 650 | 4 | |a Parasite reduction ratio | |
| 650 | 4 | |a Plasmodium falciparum | |
| 650 | 4 | |a [(3)H]hypoxanthine | |
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| 650 | 7 | |a Hypoxanthine |2 NLM | |
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| 700 | 1 | |a Niepoth, Elena |e verfasserin |4 aut | |
| 700 | 1 | |a Mavraj-Husejni, Arbreshe |e verfasserin |4 aut | |
| 700 | 1 | |a Kreidenweiss, Andrea |e verfasserin |4 aut | |
| 700 | 1 | |a Herrmann, Jennifer |e verfasserin |4 aut | |
| 700 | 1 | |a Müller, Rolf |e verfasserin |4 aut | |
| 700 | 1 | |a Knaab, Tanja |e verfasserin |4 aut | |
| 700 | 1 | |a Burckhardt, Bjoern B |e verfasserin |4 aut | |
| 700 | 1 | |a Kurz, Thomas |e verfasserin |4 aut | |
| 700 | 1 | |a Held, Jana |e verfasserin |4 aut | |
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