Sep 08, 2020
Oceanit Lateral Flow Assay (LFA) Protocol
- 1Oceanit Laboratories, Inc.
- Dexter Poon: XPrize submission
External link: https://assure19.com
Protocol Citation: proposals, Dexter Poon 2020. Oceanit Lateral Flow Assay (LFA) Protocol. protocols.io https://dx.doi.org/10.17504/protocols.io.bk3jkykn
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: In development
We are still developing and optimizing this protocol
Created: September 08, 2020
Last Modified: September 08, 2020
Protocol Integer ID: 41803
Abstract
This experiment uses infectious WA1 strain of SARS-CoV2 and is conducted in a qualified BSL3 facility at the University of Hawaii John A Burns School of Medicine (Honolulu, Hawaii).
Materials
MATERIALS
RNase A (10 mg/mL)Thermo Fisher ScientificCatalog #EN0531
Triton X-100VWR ScientificCatalog #9002-93-1
BSASigma AldrichCatalog #A2153
Gold Nanospheres - Bare (Citrate) 80nmNanocomposixCatalog # AUCN80
TBSTweenSigma AldrichCatalog #9039-10PAK
Sample PadCytivaCatalog #8134-2250
Oceanit Lateral Flow Assay (LFA) Protocol
Oceanit Lateral Flow Assay (LFA) Protocol
Prepare Serial Dilution
This experiment uses infectious WA1 strain of SARS-CoV2 and is conducted in a qualified BSL3 facility at the University of Hawaii John A Burns School of Medicine (Honolulu, Hawaii).
Aliquot diluent solution into centrifuge tubes
Diluent solution is cell growth media supplemented with 10% FBS.
Prepare lysis buffer with LFA capture molecule
Lysis buffer containing LFA capture molecule is Oceanit’s proprietary formulation.
Virus stock (4 x 10e7 pfu/mL)
Virus stock 10-fold serial dilutions.
10-fold serial dilutions and control tubes
a.Tube 1 – Neat (4e7 pfu/mL); pfu = plaque forming units
b.Tube 2 – 10e-1
c.Tube 3 – 10e-2
d.Tube 4 – 10e-3
e.Tube 5 – 10e-4
f.Tube 6 – 10e-5
g.Tube 7 – 10e-6
h.Tube 8 – 10e-7
i.Control – no virus
Add lysis buffer to dilutions
Add prepared lysis buffer to labeled individual LFA cassettes. Test and control lines are Oceanit’s proprietary molecules.
Incubate 5 minutes
Add to labeled individual LFA cassettes.
Add 250uL of the corresponding sample to individual LFA cassettes.
Record results with photography:
A.Amount:
B.Concentration: Virus stock and dilution range from 4 x 10e7 pfu/mL to 0 virus added.
C.Temperature: Virus stocks are stored at -80C; assay is performed at room temperature.
D.Duration of the experiment: 1 hour elapsed time.
E.Equipment: BSL3 containment, Class II biological safety cabinet, autoclave, pipette and disposable tips, disposable tubes, dedicated camera.
F.Reagents: Vero E6 cell culture growth media supplemented with 10% FBS, Oceanit lysis buffer + capture molecule (proprietary), LFA embedded test and control line molecules (proprietary)