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Antimicrobial Agents and Chemotherapy, January 2003, p. 170-173, Vol. 47, No. 1
0066-4804/03/$08.00+0     DOI: 10.1128/AAC.47.1.170-173.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Simple In Vitro Assay for Determining the Sensitivity of Plasmodium vivax Isolates from Fresh Human Blood to Antimalarials in Areas where P. vivax Is Endemic

Bruce M. Russell,1,2* Rachanee Udomsangpetch,3 Karl H. Rieckmann,1 Barbara M. Kotecka,1 Russell E. Coleman,2 and Jetsumon Sattabongkot2

Australian Army Malaria Institute, Enoggera QLD 4052, Australia,1 Department of Pathobiology, Faculty of Science, Mahidol University,3 Department of Entomology, U.S. Armed Forces Research Institute of Medical Sciences, Bangkok 10400, Thailand2

Received 7 June 2002/ Returned for modification 18 June 2002/ Accepted 21 October 2002

The aim of this study was to develop a simple, field-practical, and effective in vitro method for determining the sensitivity of fresh erythrocytic Plasmodium vivax isolates to a range of antimalarials. The method used is a modification of the standard World Health Organization (WHO) microtest for determination of P. falciparum drug sensitivity. The WHO method was modified by removing leukocytes and using a growth medium supplemented with AB+ serum. We successfully carried out 34 in vitro drug assays on 39 P. vivax isolates collected from the Mae Sod malaria clinic, Tak Province, Thailand. The mean percentage of parasites maturing to schizonts (six or more merozoites) in control wells was 66.5% ± 5.9% (standard deviation). This level of growth in the control wells enabled rapid microscopic determination (5 min per isolate per drug) of the MICs of chloroquine, dihydroartemisinin, WR238605 (tafenoquine), and sulfadoxine. P. vivax was relatively sensitive to chloroquine (MIC = 160 ng/ml, 50% inhibitory concentration [IC50] = 49.8 ng/ml) and dihydroartemisinin (MIC = 0.5 ng/ml, IC50 = 0.47 ng/ml). The poor response of P. vivax to both tafenoquine (MIC = 14,000 ng/ml, IC50 = 9,739 ng/ml) and sulfadoxine (MIC = 500,000 ng/ml, IC50 = 249,000 ng/ml) was due to the slow action of these drugs and the innate resistance of P. vivax to sulfadoxine. The in vitro assay developed in our study should be useful both for assessing the antimalarial sensitivity of P. vivax populations and for screening new antimalarials in the absence of long-term P. vivax cultures.

* Corresponding author. Mailing address: Department of Entomology, USAMC-AFRIMS, 315/6 Rajvithi Rd., Bangkok 10400, Thailand. Phone: 66-2-644-4888. Fax: 66-2-246-8832. E-mail: RussellBM{at}AMEDD.ARMY.MIL.

Antimicrobial Agents and Chemotherapy, January 2003, p. 170-173, Vol. 47, No. 1
0066-4804/03/$08.00+0     DOI: 10.1128/AAC.47.1.170-173.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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