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

Area of Science:

• Biotechnology
• Analytical Chemistry
• Virology

Background:

• Accurate and rapid virus detection is crucial for public health and research.
• Conventional methods like ELISA can be time-consuming and may lack sensitivity for certain applications.
• There is a need for faster, more sensitive, and field-deployable virus detection techniques.

Purpose of the Study:

• To develop a novel, rapid, and sensitive method for virus detection.
• To utilize a polyelectrolyte-multilayer-modified cellulosic filter paper for enhanced virus capture and detection.
• To compare the performance of the new method against conventional sandwich enzyme-linked immunosorbent assay (ELISA).

Main Methods:

• Development of a polyelectrolyte-multilayer-modified cellulosic filter paper assay.
• Integration of immunodetection using anti-M13 horseradish peroxidase (HRP) conjugate and 3,3',5,5'-tetramethylbenzidine (TMB) substrate.
• Testing with M13 bacteriophage as a model virus, with visual and scanner-based detection.

Main Results:

• The entire assay can be completed within 2 hours.
• Visual detection limit of 10^6 plaque-forming units (pfu)/ml with 0.50 ml sample volume.
• Improved detection limit to 5x10^4 pfu/ml by increasing sample volume to 100 ml.
• Conventional ELISA achieved a detection limit of only 10^7 pfu/ml with the same detection system.

Conclusions:

• The developed filter paper-based immunodetection assay offers a rapid and sensitive alternative for virus detection.
• The method demonstrates superior sensitivity compared to conventional ELISA, particularly when larger sample volumes are processed.
• This technique holds potential for point-of-care diagnostics and field applications requiring quick and reliable virus identification.