Enzyme-assisted upconversion fluorescence-encoded biosensing system for simultaneous detection of multiple sites EGFR...

Area of Science:

• Biomedical Engineering
• Nanotechnology
• Molecular Diagnostics

Background:

• Epidermal growth factor receptor (EGFR) mutations are critical drivers in various cancers, necessitating rapid detection for effective patient treatment.
• Accurate identification of specific EGFR mutations, such as L858R and 19Del, guides personalized therapy in non-small cell lung cancer (NSCLC).

Purpose of the Study:

• To develop a highly sensitive fluorescent biosensing system for simultaneous detection of L858R and 19Del EGFR mutations.
• To enhance signal amplification through lanthanide ion-doped upconversion nanoparticles (UCNPs) and polymerization reactions.

Main Methods:

• Preparation of two distinct UCNPs (NaYF4:Yb,Er and NaYF4:Yb,Tm) with unique fluorescence profiles.
• Signal amplification via cDNA hybridization, atom transfer radical polymerization (ATRP), and magnetic bead separation.
• Detection of mutations by analyzing characteristic upconversion fluorescence spectra after enzymatic cleavage.

Main Results:

• Simultaneous quantitative detection of L858R and 19Del mutations with a wide detection range (10 fM to 10 nM).
• Achieved ultra-low detection limits: 2.44 fM for L858R and 2.13 fM for 19Del.
• Demonstrated effective differentiation from wild-type and other mutations, with results consistent with qPCR.

Conclusions:

• The developed UCNP-based biosensing system offers high sensitivity and specificity for simultaneous EGFR mutation detection.
• This technology shows significant potential for early diagnosis and precision treatment of NSCLC.
• The system's performance supports its application in clinical diagnostics and personalized cancer therapy.