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Related Concept Videos

Enzyme-Linked Immunosorbent Assay01:33

Enzyme-Linked Immunosorbent Assay

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In 1971, Peter Perlman and Eva Engvall developed an Enzyme-linked immunosorbent assay (ELISA or EIA). ELISA differs from western blot in that the assays are conducted in microtiter plates or in vivo rather than on an absorbent membrane.
There are many different types of ELISAs, but they all involve an antibody molecule whose constant region binds an enzyme, leaving the variable region free to bind its specific antigen.  Enzyme-substrate reaction allows the antigen to be visualized or...
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Related Experiment Video

Updated: Oct 18, 2025

Zika Virus Specific Diagnostic Epitope Discovery
11:37

Zika Virus Specific Diagnostic Epitope Discovery

Published on: December 12, 2017

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Zika Virus Non-Structural Protein 1 Antigen-Capture Immunoassay.

Brandon J Beddingfield1, Jessica N Hartnett1, Russell B Wilson2

  • 1Department of Microbiology and Immunology, School of Medicine, Tulane University, New Orleans, LA 70112, USA.

Viruses
|September 28, 2021
PubMed
Summary
This summary is machine-generated.

Developing specific Zika virus (ZIKV) diagnostic tests is challenging due to genetic similarities with other flaviviruses. This study modified ZIKV NS1 protein and used affinity-purified antibodies to create a more specific antigen-capture ELISA.

Keywords:
Zika virusantigen-capture ELISAnon-structural protein 1polyclonal antibodiessite-directed mutagenesis

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Quantification of Antibody-dependent Enhancement of the Zika Virus in Primary Human Cells
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Area of Science:

  • Virology
  • Immunology
  • Diagnostic Assay Development

Background:

  • Zika virus (ZIKV) infection typically causes mild illness but can lead to severe neurological disease and major birth defects in pregnancy.
  • Genetic similarities between ZIKV and other flaviviruses, like dengue virus (DENV), complicate the development of specific diagnostic assays.
  • Nonstructural protein 1 (NS1) is a key target for flavivirus immunodiagnostic assays.

Purpose of the Study:

  • To develop a ZIKV-specific diagnostic assay by addressing cross-reactivity issues with related flaviviruses.
  • To engineer ZIKV NS1 protein variants and generate specific antibodies for improved diagnostic accuracy.

Main Methods:

  • Site-directed mutagenesis was used to modify putative epitopes on the ZIKV NS1 protein.
  • Goat polyclonal antibodies against the variant ZIKV NS1 were affinity-purified to remove cross-reactive antibodies to DENV NS1.
  • An antigen-capture ELISA was configured using the affinity-purified antibodies to detect ZIKV NS1.

Main Results:

  • The developed antigen-capture ELISA demonstrated a linear dynamic range between approximately 500 and 30 ng/mL.
  • The limit of detection for the assay was determined to be between 1.95 and 7.8 ng/mL.
  • NS1 proteins from DENV, yellow fever virus, St. Louis encephalitis virus, and West Nile virus showed significantly reduced reactivity, indicating improved ZIKV specificity.

Conclusions:

  • Modification of ZIKV NS1 and affinity purification of antibodies can significantly reduce cross-reactivity with related flaviviruses.
  • The developed ELISA shows promise for accurate ZIKV diagnosis, even in regions co-endemic with other flaviviruses.
  • Further refinement of these methods could lead to robust ZIKV-specific immunoassays for widespread clinical use.