Recent developments in high-throughput mutation screening

Area of Science:

• Pharmacogenomics
• Molecular Biology
• Genetics

Background:

• Mutation screening is crucial for pharmacogenomics.
• Automated DNA sequencing is not ideal for large-scale mutation analysis.
• High-throughput methods are needed for analyzing hundreds of thousands of mutations.

Purpose of the Study:

• To review and compare high-throughput mutation screening methods.
• To identify suitable technologies for large-scale genetic analysis.
• To discuss the potential of emerging technologies in mutation detection.

Main Methods:

• DNA microarrays and oligonucleotide chips for known mutations.
• Matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDI-TOF MS) for single nucleotide polymorphism (SNP) analysis.
• Single strand conformation polymorphism (SSCP) and conformation sensitive gel electrophoresis (CSGE) combined with capillary array electrophoresis or denaturing high-performance liquid chromatography for unknown mutations.

Main Results:

• DNA microarrays and oligonucleotide chips enable parallel hybridization for thousands of known mutations.
• MALDI-TOF MS offers fast, automatable high-throughput SNP analysis.
• SSCP/CSGE combined with electrophoresis/HPLC provide sensitive, high-throughput screening for unknown mutations with potential for further speed improvements.

Conclusions:

• Various high-throughput methods exist for mutation screening in pharmacogenomics.
• The choice of method depends on whether mutations are known or unknown.
• Advancements in capillary electrophoresis chips promise faster analysis times for sensitive mutation detection.