1. Assessment of the Worldwide Antimalarial Resistance Network Standardized Procedure for In Vitro Malaria Drug Sensitivity Testing Using SYBR Green Assay for Field Samples with Various Initial Parasitemia Levels
- Author
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Mayank Talwar, Mark Hickman, Heather Gaona, Tushar Murthy, Jacob D. Johnson, Jennifer M. Auschwitz, Edwin Kamau, Susan E. Leed, Charles O. Okello, Hoseah M. Akala, Redemptah Yeda, Agnes C. Cheruiyot, and Patricia J. Lee
- Subjects
0301 basic medicine ,Drug ,Erythrocytes ,media_common.quotation_subject ,030106 microbiology ,030231 tropical medicine ,Plasmodium falciparum ,Drug Resistance ,Parasitemia ,Diamines ,Sensitivity and Specificity ,Epidemiology and Surveillance ,03 medical and health sciences ,Antimalarials ,Inhibitory Concentration 50 ,0302 clinical medicine ,parasitic diseases ,medicine ,Humans ,Pharmacology (medical) ,Public Health Surveillance ,Benzothiazoles ,Malaria, Falciparum ,Organic Chemicals ,Atovaquone ,media_common ,Fluorescent Dyes ,Pharmacology ,biology ,Reproducibility of Results ,Chloroquine ,Drug susceptibility ,DNA, Protozoan ,biology.organism_classification ,medicine.disease ,Virology ,In vitro ,Artemisinins ,Mefloquine ,Infectious Diseases ,Drug concentration ,Sensitivity testing ,Immunology ,Quinolines ,Biological Assay ,Malaria - Abstract
The malaria SYBR green assay, which is used to profile in vitro drug susceptibility of Plasmodium falciparum , is a reliable drug screening and surveillance tool. Malaria field surveillance efforts provide isolates with various low levels of parasitemia. To be advantageous, malaria drug sensitivity assays should perform reproducibly among various starting parasitemia levels rather than at one fixed initial value. We examined the SYBR green assay standardized procedure developed by the Worldwide Antimalarial Resistance Network (WWARN) for its sensitivity and ability to accurately determine the drug concentration that inhibits parasite growth by 50% (IC 50 ) in samples with a range of initial parasitemia levels. The initial sensitivity determination of the WWARN procedure yielded a detection limit of 0.019% parasitemia. P. falciparum laboratory strains and field isolates with various levels of initial parasitemia were then subjected to a range of doses of common antimalarials. The IC 50 s were comparable for laboratory strains with between 0.0375% and 0.6% parasitemia and for field isolates with between 0.075% and 0.6% parasitemia for all drugs tested. Furthermore, assay quality (Zā²) analysis indicated that the WWARN procedure displays high robustness, allowing for drug testing of malaria field samples within the derived range of initial parasitemia. The use of the WWARN procedure should allow for the inclusion of more malaria field samples in malaria drug sensitivity screens that would have otherwise been excluded due to low initial parasitemia levels.
- Published
- 2016