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  • Filtration, adsorption and immunodetection of virus using polyelectrolyte multilayer-modified paper.

Filtration, adsorption and immunodetection of virus using polyelectrolyte multilayer-modified paper.

Colloids and surfaces. B, Biointerfaces (2012-09-27)
Per A Larsson, Sushma G Puttaswamaiah, Christine Ly, Alois Vanerek, J Christopher Hall, François Drolet
ABSTRACT

A new method for detection of viruses has been developed. The entire assay can be performed within 2h, and consists of a polyelectrolyte-multilayer-modified cellulosic filter paper combined with immunodetection. The M13 bacteriophage was used as a model virus. A visual colour-based detection system, anti-M13 horseradish peroxidase (HRP) conjugate and 3,3',5,5'-tetramethylbenzidine (TMB), was selected to allow semi-quantitative assessment by human eye, or quantitative assessment using a digital scanner. By filtering a volume of 0.50 ml, it was possible to visually detect a concentration of 10(6) pfu/ml. The detection limit was improved to 5×10(4) pfu/ml by increasing the volume of the sample to 100ml. For comparison, it was only possible to detect a concentration of 10(7) pfu/ml using conventional sandwich enzyme-linked immunosorbent assay (ELISA) with the same detection system.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
3,3′,5,5′-Tetramethylbenzidine, ≥98.0% (NT)
Supelco
3,3′,5,5′-Tetramethylbenzidine, standard for GC
Sigma-Aldrich
3,3′,5,5′-Tetramethylbenzidine, ≥99%
Sigma-Aldrich
3,3′,5,5′-Tetramethylbenzidine, ≥98% (TLC)
Sigma-Aldrich
3,3′,5,5′-Tetramethylbenzidine, tablet, 1 mg substrate per tablet
Sigma-Aldrich
Poly(allylamine hydrochloride), average Mw ~17,500 (GPC vs. PEG std.)
Sigma-Aldrich
Poly(allylamine hydrochloride), average Mw 50,000
Sigma-Aldrich
Poly(allylamine) solution, 20 wt. % in H2O
Sigma-Aldrich
Poly(allylamine) solution, average Mw ~65,000, 10 wt. % in H2O