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Table 1 Comparison of RSA protocols

From: Alternative methods for the Plasmodium falciparum artemisinin ring-stage survival assay with increased simplicity and parasite stage-specificity

Variables

Percoll gradient RSA [8]

Sorbitol-only RSA

Filtration RSA

Chemicals required during RSAa

Heparin, percoll, sorbitol

Sorbitol

None

Volume of parasite culture needed

≥300 µL packed RBCs at 2 % parasitemia

40 µL packed RBCs at 1 % parasitemia

≥1.5 mL packed RBCs at 2–3 % parasitemia

Number of steps until completion

17

10

12

Preparation time, before drug pulse

4.5 h

30 min

3 h

Total time of parasite incubation from drug pulse

66 h

66 h

66 or 90 h, if DMSO <0.40 % at 66 h

Stage distribution at time of drug pulse

80 % rings, 20 % trophozoites and schizonts

Mixture of young and older rings

3 h rings only

Presence of pyknotic parasites in read out

Yes

Yes

No

  1. aAll methods require sorbitol synchronization two parasite cycles before performing RSA