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Table 1 Details of SMFA methods

From: An inter-laboratory comparison of standard membrane-feeding assays for evaluation of malaria transmission-blocking vaccines

Parameters LMVR TropIQ
All tests mAbs Human pAb
Gametocyte culture
 Flask 25 or 75 cm2 flask Tipper culture flask
 Htca (%) 5.0 5.0 5.0
Feeding
 Total volume (μL) 260 270 1200
 Liquid volume (μL) 160 119 696
 Liquid contents 100 μL of human serum plus 60 μL of sample in 1xPBS 119 μL of human serumb 696 μL of human serumb
 Htcc (%) 38.5 55.9 42.0
 Stage V gametocyted (%) 0.1–0.2 0.1–1.0 NDe
 Starving mosquitoes (h) 18–21 1 1
 Time of feed (min) 20 10 10
 Elimination of unfed mosquitoes from analysis Only mosquitoes with eggs at the time of dissection are analysed One hour after feeding, fully fed mosquitoes are selected transported to a bigger cage
Dissection
 Days post feed (day) 8 8 8
 Number of mosquitoes 20 20 30
Quality control
 Pre-feeding >0.5 % of stage V gametocytaemia and >800/mL of exflagellating centres in the undiluted culture Positive for male exflagellating gametocytes
 Post-feeding >4 oocysts/midgut in the assay controls >70 % infected mosquitoes in the assay controls
  1. aHaematocrit
  2. bAdd lyophilized human serum to make the liquid volume for test antibody to be 100 % human serum
  3. cFinal haematocrit in a feeder
  4. dFinal stage V gametocytaemia in a feeder
  5. eNot determined