We present a novel method to monitor DNA amplification and to analyze the dissociation curve of the amplicons using fiber optic surface plasmon resonance (SPR) sensors. Our unique SPR fibers can be combined with any benchtop thermocycler. A key factor to enable sub-nanomolar DNA detection is the efficient labeling of the reverse primers with gold-nanoparticles (AuNP). The sharp melting profiles of the AuNP-labeled amplicons allow for the detection and the location of single point mutations (SNPs). We obtained data of SNP detection in DNA targets up to 80bp long and compared these results with standard high resolution melting curve (HRM) analysis to prove that the accuracy of the sensor is as good as or, in some applications, better than the current fluorescent based systems. Because no fluorescent reporters are involved, we can screen for DNA markers in opaque media which opens perspectives for PCR analysis of less diluted complex samples.

Fiber optic SPR assay for enhanced real-time PCR analysis of non-transparent samples

243rd National Meeting (2012)

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