Table 4.
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.