Paper-based Vertical Flow Immunoassay (VFI) for detection of bio-threat pathogens

Peng Chen, Marcellene Gates-Hollingsworth, Sujata Pandit, Anson Park, Douglas Montgomery, David AuCoin, Jian Gu, Frederic Zenhausern

Research output: Contribution to journalArticle

Abstract

Currently, the standard method for identifying biological agents of potential threats to national security and public health, such as pathogens, virus, and toxins, mainly rely on microbiological cultivation. This method is time-consuming and it requires sophisticated equipment and well-trained personnel, which are often unavailable in remote areas or at point-of-need. Therefore, an alternative rapid, simple, and sensitive method for detecting bio-threat agents is in crucial need. We report a paper-based Vertical Flow Immunoassay (VFI) device that can overcome these limitations. The VFI device utilizes a nanoporous nitrocellulose membrane encapsulated in a stainless steel filter holder. As the sample is pushed through the membrane, which is pre-functionalized with capture antibody, a sandwich assay is formed and colorimetric signal is generated to reflect the presence of target antigens. Through theoretical analyses of antigen-antibody binding process inside a porous membrane, we identified two critical factors – membrane pore size and sample flow rate that can be optimized to improve the assay sensitivity. Then, the effects were demonstrated through experimental studies using Burkholderia pseudomallei (the causative agent of melioidosis) as a model pathogen. The B. pseudomallei VFI was based on an immunoassay targeting the B. pseudomallei surface capsular polysaccharide (CPS). The experimental results agreed well with the theory showing that increasing the flow speed (up to 1.06 mm/s) and reducing the membrane pore size (down to 0.1 µm) could improve the sensitivity by at least 5 times. The VFI's limit-of-detection for CPS spiked in buffer solution was determined to be 0.02 ng/mL. The developed VFI shows great potential as a point-of-care tool for detection of bio-threat agents in a variety of clinical and resource-restricted conditions.

LanguageEnglish (US)
Pages81-88
Number of pages8
JournalTalanta
Volume191
DOIs
StatePublished - Jan 1 2019
Externally publishedYes

Fingerprint

Pathogens
Immunoassay
Burkholderia pseudomallei
Membranes
Equipment and Supplies
Pore size
Polysaccharides
Assays
Point-of-Care Systems
Security Measures
Melioidosis
Antigens
Collodion
National security
Antibodies
Stainless Steel
Biological Factors
Public health
Viruses
Sample Size

Keywords

  • Biological threat reduction
  • Burkholderia pseudomallei
  • Melioidosis
  • Paper-based microfluidic
  • Point-of-care diagnostic
  • Vertical Flow Immunoassay

ASJC Scopus subject areas

  • Chemistry(all)

Cite this

Chen, P., Gates-Hollingsworth, M., Pandit, S., Park, A., Montgomery, D., AuCoin, D., ... Zenhausern, F. (2019). Paper-based Vertical Flow Immunoassay (VFI) for detection of bio-threat pathogens. Talanta, 191, 81-88. https://doi.org/10.1016/j.talanta.2018.08.043

Paper-based Vertical Flow Immunoassay (VFI) for detection of bio-threat pathogens. / Chen, Peng; Gates-Hollingsworth, Marcellene; Pandit, Sujata; Park, Anson; Montgomery, Douglas; AuCoin, David; Gu, Jian; Zenhausern, Frederic.

In: Talanta, Vol. 191, 01.01.2019, p. 81-88.

Research output: Contribution to journalArticle

Chen, P, Gates-Hollingsworth, M, Pandit, S, Park, A, Montgomery, D, AuCoin, D, Gu, J & Zenhausern, F 2019, 'Paper-based Vertical Flow Immunoassay (VFI) for detection of bio-threat pathogens' Talanta, vol. 191, pp. 81-88. https://doi.org/10.1016/j.talanta.2018.08.043
Chen, Peng ; Gates-Hollingsworth, Marcellene ; Pandit, Sujata ; Park, Anson ; Montgomery, Douglas ; AuCoin, David ; Gu, Jian ; Zenhausern, Frederic. / Paper-based Vertical Flow Immunoassay (VFI) for detection of bio-threat pathogens. In: Talanta. 2019 ; Vol. 191. pp. 81-88.
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