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Related Concept Videos

Enzyme-Linked Immunosorbent Assay01:33

Enzyme-Linked Immunosorbent Assay

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 quantified.
Immunoprecipitation01:20

Immunoprecipitation

Immunoprecipitation, or IP, is a widely used technique that employs protein-antibody interactions to isolate proteins or protein complexes in their native state for studying protein-protein interactions, quaternary structures, or supramolecular complexes. Various modifications of the technique, including chromatin IP, cross-linking IP, and fluorescence IP, are commonly used.
Chromatin Immunoprecipitation
Chromatin immunoprecipitation, also known as ChIP, is used to study protein-DNA or...

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Antigen-Capture Enzyme-Linked Immunosorbent Assay for Specific Detection of Mycoplasma pneumoniae
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Antigen-Capture Enzyme-Linked Immunosorbent Assay for Specific Detection of Mycoplasma pneumoniae

Published on: February 24, 2023

Sample handling substantially affects Johne's ELISA.

Catherine A Alinovi1, Michael P Ward, Tsang Long Lin

  • 1Department of Comparative Pathobiology, School of Veterinary Medicine, Purdue University, West Lafayette, IN 47907-2065, USA.

Preventive Veterinary Medicine
|May 30, 2009
PubMed
Summary
This summary is machine-generated.

Sample handling and storage significantly impact Mycobacterium avium subsp. paratuberculosis (MAP) ELISA test results for Johne's disease diagnosis. Proper procedures are essential for accurate cattle classification and disease control.

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High Throughput Sequential ELISA for Validation of Biomarkers of Acute Graft-Versus-Host Disease

Published on: October 31, 2012

Area of Science:

  • Veterinary Diagnostics
  • Immunology
  • Bacteriology

Background:

  • Accurate detection of Mycobacterium avium subsp. paratuberculosis (MAP) is critical for diagnosing Johne's disease in cattle.
  • Current diagnostic methods, including ELISA, have limitations that can affect reliability.
  • Variability in sample handling and storage can introduce confounding factors into diagnostic test results.

Purpose of the Study:

  • To evaluate the impact of different transportation, preparation, and storage conditions on MAP ELISA results.
  • To identify specific pre-analytical variables that significantly alter Johne's ELISA scores.
  • To assess how these changes affect the accurate classification of MAP-infected cattle.

Main Methods:

  • Blood samples from 21 cows with varying MAP ELISA scores were used.
  • Samples underwent different transportation temperatures (ice, 26°C) and preparation methods (serum, hemolyzed, clotted whole blood).
  • Storage conditions (4°C, -20°C for one week) and processing times (1 day, 1 week) were also tested.

Main Results:

  • Higher MAP ELISA scores were observed for whole blood and hemolyzed samples transported at 26°C compared to serum-separated samples (P=0.027).
  • Storage for one week at -20°C significantly decreased MAP ELISA scores compared to 4°C storage (P<0.001).
  • Both sample preparation and storage temperature demonstrably affected ELISA outcomes.

Conclusions:

  • Sample preparation methods, transportation temperature, and storage temperature significantly influence MAP ELISA results.
  • These pre-analytical variations can lead to misclassification of infected cattle.
  • Standardized protocols are necessary to ensure accurate diagnosis and effective Johne's disease control programs.