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Related Concept Videos

Enzyme-Linked Immunosorbent Assay

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.

Real Time RT-PCR

Real Time RT-PCR

Real-time reverse transcription-polymerase chain reaction, or Real-time RT-PCR, is an analytical tool used to determine the expression level of target genes. The method involves converting mRNA to complementary DNA with the help of an enzyme known as reverse transcriptase, followed by the PCR amplification of the cDNA. These two processes can be performed simultaneously in a single tube or separately as a two-step reaction.
The real-time quantification of the number of amplified products is...

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Related Experiment Video

Updated: Jun 4, 2026

Amplicon Sequencing using the Long-Read Sequencing Technologies

Amplicon Sequencing using the Long-Read Sequencing Technologies

Published on: August 29, 2025

Quantifying amplicons with ELISA.

O Lantz¹, E Bonney, Y Taoufik

¹Inserm Unit, Necker Hospital, Paris, France.

Methods in Molecular Medicine

|February 23, 2011

Summary

This summary is machine-generated.

Enzyme-linked immunosorbent assay (ELISA) provides a sensitive, specific, and versatile method for quantifying polymerase chain reaction (PCR) products. Luminometry-based ELISA offers the widest dynamic range and highest sensitivity for accurate nucleic acid quantification.

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Area of Science:

Molecular Biology
Biochemistry
Assay Development

Background:

Quantitative polymerase chain reaction (PCR) requires sensitive and specific methods for measuring PCR product amounts.
Existing assays for nucleic acid quantification often lack versatility, dynamic range, or ease of use.
Enzyme-linked immunosorbent assay (ELISA) presents a promising alternative for accurate amplicon quantification.

Purpose of the Study:

To evaluate the suitability of enzyme-linked immunosorbent assay (ELISA) for quantifying polymerase chain reaction (PCR) products.
To explore different detection methods within ELISA, including colorimetry, fluorometry, and luminometry.
To highlight the advantages of ELISA, particularly luminometry, for quantitative PCR applications.

Main Methods:

Utilizing enzyme-linked immunosorbent assay (ELISA) for the quantification of polymerase chain reaction (PCR) amplicons.
Comparing detection methods: colorimetry, fluorometry, and luminometry.
Implementing ELISA in kinetic quantitative PCR and with internal standards.

Main Results:

ELISA-based quantification of PCR products meets key criteria: sensitivity, specificity, wide dynamic range, non-radioactivity, ease of use, and cost-effectiveness.
Luminometry-based ELISA demonstrates superior sensitivity and the broadest dynamic range among the evaluated methods.
The described ELISA format is automatable and compatible with existing laboratory equipment.

Conclusions:

Enzyme-linked immunosorbent assay (ELISA) is a highly effective method for quantifying PCR products, offering significant advantages over traditional assays.
Luminometry-based ELISA provides the most sensitive and dynamic range for accurate nucleic acid quantification.
This ELISA approach is suitable for various quantitative PCR applications, including kinetic analysis and internal standard-based quantification.