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. Author manuscript; available in PMC: 2022 Sep 2.
Published in final edited form as: J Virol Methods. 2021 Aug 9;297:114262. doi: 10.1016/j.jviromet.2021.114262

Table 4.

Inactivation of WPV1-spiked stool suspensions in blind passage.

Number of CPE Positive Flasks
Extraction Buffer Spiked Suspension:Buffer Ratio Additional Components Added Incubation time Passage 1 Passage 2
PBS (control) 1:4 NA 30 min 6/6 6/6
Trizol LS 1:3 None 5 min 0/3 0/3
Viral RNA Buffer 1:3 0.5% β-mercaptoethanol 5 min 0/3 0/3
Buffer AVL 1:4 Ethanol (44 % final) 5 min 0/3 0/3
UNEX 1:1 Ethanol (50 % final) 10 min 0/3 0/3
Binding Buffer 1:1 Proteinase K 10 min (with Pro K) 0/3 0/3
MagMAX Lysis/Binding Concentrate 1:2 None 15 min 0/3 0/3
4 M Guanidine Thiocyanate+ 20 % Ethanol 1:4 Ethanol (20 % final) 30 min 6/6 NT
4 M Guanidine Thiocyanate+ 30 % Ethanol 1:4 Ethanol (30 % final) 30 min 0/3 0/3
4 M Guanidine Thiocyanate+ 50 % Ethanol 1:4 Ethanol (50 % final) 30 min 0/3 0/3

WPV1-positive stool suspensions were incubated at room temperature with extraction buffers for the amount of time indicated. Samples were concentrated by ultrafiltration and incubated with RD cells. CPE was scored after 5 days for both 1st and 2nd passages. NT, Not Tested.