Detection of TP53 mutation using a portable surface plasmon resonance DNA-based biosensor.

Area of Science:

• Biomedical Engineering
• Molecular Biology
• Genetics

Background:

• TP53 gene mutations are critical in cancer development.
• Accurate and sensitive detection methods for TP53 mutations are essential for diagnosis and treatment.
• Existing methods can be complex or expensive.

Purpose of the Study:

• To develop and validate a DNA-based surface plasmon resonance (SPR) biosensor for detecting TP53 mutations.
• To assess the sensor's analytical performance, including selectivity, sensitivity, and reproducibility.
• To apply the biosensor to real-world samples from cell lines with and without TP53 mutations.

Main Methods:

• Development of a DNA-based SPR biosensor using thiol-derivatised oligonucleotide probes immobilized on gold surfaces.
• Optimization of probe immobilization and hybridization reactions.
• Testing with synthetic DNA sequences (complementary, non-complementary, and single-point mutation) and PCR-amplified DNA from TP53 mutated and wild-type cell lines.
• Analysis of sensor parameters: selectivity, sensitivity, reproducibility, and analysis time.

Main Results:

• The DNA-based SPR biosensor demonstrated high selectivity, distinguishing between DNA sequences with only a single base difference.
• The sensor system showed good sensitivity and reproducibility in detecting TP53 mutations.
• Successful application to real samples from Jurkat (wild-type) and Molt 4 (mutated) cell lines, accurately identifying the TP53 mutation at codon 248.
• The analysis time was efficient, making it suitable for practical applications.

Conclusions:

• The developed DNA-based SPR biosensor is a sensitive and selective tool for detecting TP53 gene mutations.
• The biosensor system shows significant potential as a cost-effective and rapid method for genetic mutation analysis.
• This technique offers a strong candidate for the routine detection of gene mutations in clinical and research settings.