Abstract
We proposed a coding and decoding method of suspension array (SA) based on micro-quartz pieces (MQPs) with different optical thicknesses. The capture probes (cDNA) were grafted onto the surfaces of MQPs and specifically recognized and combined with the partial sequence of the target DNA (tDNA) to form a MQP-cDNA-tDNA complex. Quantum dot-labeled signal probes were then used to specifically recognize and bind another portion of the tDNA in the complex to form a double-probe sandwich structure. This optical thickness-encoded SA can be decoded and detected by a dual-wavelength digital holographic phase fluorescence microscope system. We conducted a series of DNA molecule detection experiments by using this encoding method. Control experiments confirmed the specificity of optical thickness-encoded SA in DNA detection. The concentration gradient experiments then demonstrated the response of the MQPs based SA to analyte concentration. Finally, we used the encoding method to detect three types of DNA in a single sample and confirmed the feasibility of the proposed optical thickness-encoded SA in multiplexed DNA detection. The detection results are stable, and the detection exhibits high specificity and good repeatability.
Highlights
As disease diagnosis and treatment progresses, the demand for the high-throughput and multiplex analysis of numerous biomolecules within a single sample has increased
We proposed a novel multiplexed DNA detection method
micro-quartz pieces (MQPs) with different optical thicknesses are used as microcarriers; the optical thickness of MQPs is used as the encoding signal, and the fluorescence on the microcarriers; the optical thickness of MQPs is used as the encoding signal, and the fluorescence on signal probe is used as the label signal
Summary
As disease diagnosis and treatment progresses, the demand for the high-throughput and multiplex analysis of numerous biomolecules within a single sample has increased. Most of these diseases, including thalassemia, HPV infectious diseases, uveitis and HIV, are caused by gene deletion, gene translocation, gene mutation, or protein variation [1,2,3]. Many types of these diseases exist, but the quality of diagnosis of these diseases is limited by small sample volumes. The preparation process of SA is simple, and the detection result is stable
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