Detection of mutations: from Ames test to duplex sequencing

Area of Science:

• Genetics and Molecular Biology
• Toxicology and Mutagenesis Research

Background:

• Mutation is a fundamental biological process across all life forms.
• Detecting mutagens became crucial following the discovery of chemical mutagens in the 1940s.
• Early mutagenicity studies utilized organisms like Escherichia coli and Drosophila.

Purpose of the Study:

• To review the historical development of mutagenicity testing.
• To explore the genetic basis of the Ames test.
• To highlight Duplex Sequencing (DS) as an emerging tool for sensitive mutation detection.

Main Methods:

• Review of historical mutagenicity testing methodologies.
• Discussion of bacterial assays, including the Ames test.
• Introduction to Next-Generation Sequencing (NGS) and error-corrected NGS (ecNGS) techniques, specifically Duplex Sequencing (DS).

Main Results:

• The Ames test revolutionized mutagenicity screening by using bacteria to detect DNA-mutating capabilities of chemicals.
• Sanger sequencing provided detailed DNA sequence change information.
• Next-Generation Sequencing (NGS) and ecNGS techniques, like DS, enable large-scale, sensitive mutation analysis without phenotypic selection.

Conclusions:

• The Ames test remains a widely adopted assay for mutagenicity testing.
• Duplex Sequencing (DS) represents a significant advancement in detecting low-frequency mutations.
• These evolving techniques are crucial for understanding mutation processes and identifying harmful mutagens.