Optimizing the HRP-2 in vitro malaria drug susceptibility assay using a reference clone to improve comparisons of Plasmodium falciparum field isolates
| Autor: | Duong Socheat, Douglas S. Walsh, Delia Bethell, Paktiya Teja-Isavadharm, Harald Noedl, Kritsanai Yingyuen, Youry Se, Panjaporn Chaichana, David L. Saunders, Kurt Schaecher, Stuart D. Tyner, Chanthap Lon, Mark M Fukuda, Suwanna Chaorattanakawee, Wiriya Rutvisuttinunt |
|---|---|
| Rok vydání: | 2012 |
| Předmět: |
Plasmodium falciparum
lcsh:Arctic medicine. Tropical medicine lcsh:RC955-962 medicine.medical_treatment Clone (cell biology) Protozoan Proteins Dihydroartemisinin Artesunate Anti-malarial drugs Antigens Protozoan HRP-2 Drug resistance Pharmacology Mass Spectrometry lcsh:Infectious and parasitic diseases Microbiology chemistry.chemical_compound Antimalarials Inhibitory Concentration 50 Parasitic Sensitivity Tests parasitic diseases medicine Humans lcsh:RC109-216 Artemisinin Malaria Falciparum biology Research biology.organism_classification medicine.disease Artemisinins Culture Media Malaria Infectious Diseases Parasitology chemistry Drug susceptibility test ELISA medicine.drug Chromatography Liquid |
| Zdroj: | Malaria Journal Malaria Journal, Vol 11, Iss 1, p 325 (2012) |
| ISSN: | 1475-2875 |
| Popis: | Background Apparent emerging artemisinin-resistant Plasmodium falciparum malaria in Southeast Asia requires development of practical tools to monitor for resistant parasites. Although in vitro anti-malarial susceptibility tests are widely used, uncertainties remain regarding interpretation of P. falciparum field isolate values. Methods Performance parameters of the W2 P. falciparum clone (considered artemisinin “sensitive”) were evaluated as a reference for the HRP-2 immediate ex vivo assay. Variability in W2 IC50s was assessed, including intra- and inter-assay variability among and between technicians in multiple experiments, over five freeze-thaw cycles, over five months of continuous culture, and before and after transport of drug-coated plates to remote field sites. Nominal drug plate concentrations of artesunate (AS) and dihydroartemisinin (DHA) were verified by LC-MS analysis. Plasmodium falciparum field isolate IC50s for DHA from subjects in an artemisinin-resistant area in Cambodia were compared with W2 susceptibility. Results Plate drug concentrations and day-to-day technical assay performance among technicians were important sources of variability for W2 IC50s within and between assays. Freeze-thaw cycles, long-term continuous culture, and transport to and from remote sites had less influence. Despite variability in W2 susceptibility, the median IC50s for DHA for Cambodian field isolates were higher (p Conclusion The W2 reference clone improved the interpretability of field isolate susceptibility from the immediate ex vivo HRP-2 assay from areas of artemisinin resistance. Methods to increase the reproducibility of plate coating may improve overall assay interpretability and utility. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
Full text from SpringerLink