AuNPs beacons-enhanced surface plasmon resonance imaging sensor for rapid, high-throughput and ultra-sensitive detection of three fusion genes related to acute promyelocytic leukemia

Abstract

In this study, an ultra-sensitive, AuNPs beacons-enhanced plasma sensor for detecting three isoforms of PML/RARA fusion gene was developed. The detection and classification of three subtypes of PML/RARA genomic fragments is of significance for the prediction, risk stratification and precise drug administration in acute promyelocytic leukemia (APL). Over here, SPRi sensor applied freezing method to immobilize the probe, thus the time required for probe immobilization to be reduced from the commonly used 24 h incubation to 2 h, which greatly improves the detection efficiency. The nanoparticle-enhanced sandwich structure selected for signal amplification was constructed by a one-step method, and the nanoparticle plasma approach combining coupling and mass effects holds greater potential for signal amplification. In this system, the AuNPs beacons-enhanced SPRi established by reducing the distance between the AuNPs and the bare gold chip to improve the “coupling” effect has proved to be two orders of magnitude higher in the detection limit compared with the conventional method. The enhanced SPRi sensor is used to detect PML/RARA L isoforms, the detection limit is down to 15.57 fM with a wide linear range from 40fM to 100 pM. Furthermore, it has been proved that three isoforms of APL related fusion genes can be quantified in the high-throughput SPRi sensor while showing satisfactory specificity. Herein, the detection sensor in this paper presents a simple, time-saving, high-sensitive and high-throughput approach for the detection of fusion genes, and presents a rapid paradigm that can be easily generalized for APL diagnosis.