Paper-based Detection of Epstein-Barr Virus Using Asymmetric Polymerase Chain Reaction and Gold Silicon Particles

A new paper-based lateral flow nucleic acid (LFNA) test platform was established in this study using asymmetric polymerase chain ceaction (A-PCR) for signal amplification. This new method allowed a visual detection of Epstein-Barr virus(EBV) nucleic acids with high specificity and low cost. In addition, as part of our strategy we employed a sandwich system of capture probe (CP) / gold nanoparticles (AuNPS) and silicon dioxide (SiO2) (AuNPS@SiO2) nanospheres/target DNA/avidin complexes as the sensing platform. Biotin-labeled target DNA was obtained by A-PCR and later introduced in the LF device. The CP/target DNA/AuNPS@SiO2 complexes were captured on the test zone by the specific reaction between biotin and avidin, and the remaining CP/AuNPS@SiO2 particles were captured on the quality control zone by the hybridization between CP and a quality control probe. Au@SiO2 accumulation in the test and quality control zones of the device enabled a visual detection of the specific target sequences. The method detection limit was 50 nM of the target DNA, which was lower than that of the LFNA biosensor ( LFNAB) without PCR amplification and Au@SiO2 particles. In conclusion, the novel paper-based platform described here is a low cost, efficient and fast visual detection method that offers high sensitivity and other benefits compared to alternative methods in use.

Automated summary

A new paper-based lateral flow nucleic acid test platform using asymmetric polymerase chain reaction for signal amplification was developed, enabling visual detection of Epstein-Barr virus nucleic acids with high specificity and low cost.