The online version of this article (doi:10.1186/1475-2875-11-272) contains supplementary material, which is available to authorized users.
The authors declare that they have no competing interests.
NTH and KK developed the idea for the project. TTM and NTH conceived and designed the experiments. TTM, NTH and MNS carried out the laboratory work. TTM, NTH, DTXT, KH, and KK analysed and interpreted the data. TTM, NTH, MNS, KH and KK contributed reagents/materials/analysis tools. . TTM, NTH, DTXT and KK wrote the paper. All authors had full access to all data in the study, read and approved the manuscript.
The spread of drug resistance in malaria parasites and the limited number of effective drugs for treatment indicates the need for new anti-malarial compounds. Current assays evaluating drugs against Plasmodium falciparum require expensive materials and equipment, thus limiting the search for new drugs, particularly in developing countries. This study describes an inexpensive procedure that is based on the advantage of a positive correlation between the haemozoin level of infected erythrocytes and parasite load.
The relationship between parasitaemia and the haemozoin level of infected erythrocytes was investigated after converting haemozoin into monomeric haem. The 50% inhibitory concentration (IC50) values of chloroquine, quinine, artemisinin, quinidine and clotrimazole against P. falciparum K1 and 9A strains were determined using the novel assay method.
The haemozoin of parasites was extracted and converted into monomeric haem, allowing the use of a colorimeter to efficiently and rapidly measure the growth of the parasites. There was a strong and direct linear relationship between the absorbance of haem converted from haemozoin and the percentage of the parasite (R2 = 0.9929). Furthermore, the IC50 values of drugs were within the range of the values previously reported.
The haemozoin-based colorimetric assay can be considered as an alternative, simple, robust, inexpensive and convenient method, making it applicable in developing countries.
Breman JG, Alilio MS, Mills A: Conquering the intolerable burden of malaria: what's new, what's needed: a summary. AmJTrop Med Hyg. 2004, 71: 1-15.
WHO: Guidelines for the treatment of malaria. 2006, World Health Organization, Geneva, Switzerland
Dondorp AM, Nosten F, Yi P, Das D, Phyo AP, Tarning J, Lwin KM, Ariey F, Hanpithakpong W, Lee SJ, Ringwald P, Silamut K, Imwong M, Chotivanich K, Lim P, Herdman T, An SS, Yeung S, Singhasivanon P, Day NP, Lindegardh N, Socheat D, White NJ: Artemisinin resistance in Plasmodium falciparum malaria. N Engl J Med. 2009, 361: 455-467. 10.1056/NEJMoa0808859. PubMedCentralCrossRefPubMed
Noedl H, Se Y, Sriwichai S, Schaecher K, Teja-Isavadharm P, Smith B, Rutvisuttinunt W, Bethell D, Surasri S, Fukuda MM, Socheat D, Chan Thap L: Artemisinin resistance in Cambodia: a clinical trial designed to address an emerging problem in Southeast Asia. Clin Infect Dis. 2010, 51: e82-e89. 10.1086/657120. CrossRefPubMed
Corbett Y, Herrera L, Gonzalez J, Cubilla L, Capson TL, Coley PD, Kursar TA, Romero LI, Ortega-Barria E: A novel DNA-based microfluorimetric method to evaluate antimalarial drug activity. AmJTrop Med Hyg. 2004, 70: 119-124.
Vossen MG, Pferschy S, Chiba P, Noedl H: The SYBR Green I malaria drug sensitivity assay: performance in low parasitemia samples. AmJTrop Med Hyg. 2010, 82: 398-401. CrossRef
Makler MT, Ries JM, Williams JA, Bancroft JE, Piper RC, Gibbins BL, Hinrichs DJ: Parasite lactate dehydrogenase as an assay for Plasmodium falciparum drug sensitivity. AmJTrop Med Hyg. 1993, 48: 739-741.
Druilhe P, Moreno A, Blanc C, Brasseur PH, Jacquier P: A colorimetric in vitro drug sensitivity assay for Plasmodium falciparum based on a highly sensitive double-site lactate dehydrogenase antigen-capture enzyme-linked immunosorbent assay. AmJTrop Med Hyg. 2001, 64: 233-241.
Eckman JR, Modler S, Eaton JW, Berger E, Engel RR: Host heme catabolism in drug-sensitive and drug-resistant malaria. J Lab Clin Med. 1977, 90: 767-770. PubMed
Maeno Y, Nakazawa S, le Dao D, Yamamoto N, Giang ND, Van Hanh T, le Thuan K, Taniguchi K: A dried blood sample on filter paper is suitable for detecting Plasmodium falciparum gametocytes by reverse transcription polymerase chain reaction. Acta Trop. 2008, 107: 121-127. 10.1016/j.actatropica.2008.05.001. CrossRefPubMed
Takeuchi Y, Yamakawa N, Kaetsu M, Fujiwara K, Okada J: Experiences with AnaeroPack systems. J Jpn Assoc Anaerob Infect Res. 1992, 22: 106-112.
Bhattacharya A, Mishra LC, Bhasin VK: In vitro activity of artemisinin in combination with clotrimazole or heat-treated amphotericin B against Plasmodium falciparum. AmJTrop Med Hyg. 2008, 78: 721-728.
Kurosawa Y, Dorn A, Kitsuji-Shirane M, Shimada H, Satoh T, Matile H, Hofheinz W, Masciadri R, Kansy M, Ridley RG: Hematin polymerization assay as a high-throughput screen for identification of new antimalarial pharmacophores. Antimicrob Agents Chemother. 2000, 44: 2638-2644. 10.1128/AAC.44.10.2638-2644.2000. PubMedCentralCrossRefPubMed
Franke-Fayard B, Djokovic D, Dooren MW, Ramesar J, Waters AP, Falade MO, Kranendonk M, Martinelli A, Cravo P, Janse CJ: Simple and sensitive antimalarial drug screening in vitro and in vivo using transgenic luciferase expressing Plasmodium berghei parasites. Int J Parasitol. 2008, 38: 1651-1662. 10.1016/j.ijpara.2008.05.012. CrossRefPubMed
- A simple and inexpensive haemozoin-based colorimetric method to evaluate anti-malarial drug activity
Tran Thanh Men
Nguyen Tien Huy
Dai Thi Xuan Trang
Mohammed Nasir Shuaibu
- BioMed Central
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