Novel Signal-Enhancing Approaches for Optical Detection of Nucleic Acids—Going beyond Target Amplification
Abstract
:1. Introduction
2. Enzyme Labeling Strategies without Polymerase Target Amplification
3. Enzyme-Free Hybridization Assays: Specificity in Focus
4. Ultra-Bright Fluorophores for Nucleic Acid Labeling: Macrofluorophores, Nucleic Acid Binding Dyes, and Nanoparticles
5. Signal Enhancement in Surface Plasmon Resonance Assays
6. Signal Enhancement with Optical Detectors
Assay/Producer | Apparatus Specifications: | Compatible Labels; Absorbance-/Emission (abs-em) Range, nm; QY | LOD * | |||
---|---|---|---|---|---|---|
Light Source | Detector | Application | pmol | Ref. | ||
Colorimetry: Konica Minolta/Hach | Filament lamp or LED | PR | Chromophore with abs at 380–780 nm | DNA | 5 × 10−3 | [50,57] |
Fluorometry: Promega Glomax | Xe lamp or LED | PD/EPC + PMT | Fluorophore with abs-em at 260–750 nm, QY > 0.1 | Luciferase | 3 × 10−9 | [58] |
Horiba spectrofluorometer | Xe lamp | PD/EPC + PMT | Fluorophore with abs-em at 185–1700 nm, QY > 0.1 | DNA | FLS: 9 × 10−4 LSC: 2 × 10−4 | [59] |
TECAN multimode reader: Infinite F200 PRO | Xe lamp | PD/EPC + PMT | Fluorophore with abs-em at 230–1000 nm, QY > 0.1 | DNA | FLS: 3 × 10−4LSC: 1.2 × 10−5 | [60] |
RT-qPCR: Roche Light Cycler 480 | HI LED | CCD | Fluorophore with abs-em at 488–660 nm, QY > 0.1 | DNA | 3 × 10−3 | [61] |
Microscopy: Nikon/Leica/Olympus/Zeiss/Horiba | Lamp (Xe, MH)/LED | PD/EPC + PMT; CCD, CID, CMOS, EBCCD; GaAsP | Photostable fluorophores with abs-em at 240–940 nm, optimal at QY > 0.6 | DNA/RNA | <1 × 10−13 | [62] |
Surface plasmon resonance: Biacore, BioNavis | Lamp, diode, laser or fiber | PD or CCD | n/a | DNA | 3 × 10−4 | [64] |
7. Conclusion and Perspective: Moving toward Sensitive, Specific and Detailed Nucleic Acid Analysis Using Non-PCR Assays
Acknowledgments
Conflicts of Interest
References
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Miotke, L.; Barducci, M.C.; Astakhova, K. Novel Signal-Enhancing Approaches for Optical Detection of Nucleic Acids—Going beyond Target Amplification. Chemosensors 2015, 3, 224-240. https://doi.org/10.3390/chemosensors3030224
Miotke L, Barducci MC, Astakhova K. Novel Signal-Enhancing Approaches for Optical Detection of Nucleic Acids—Going beyond Target Amplification. Chemosensors. 2015; 3(3):224-240. https://doi.org/10.3390/chemosensors3030224
Chicago/Turabian StyleMiotke, Laura, Maria Carla Barducci, and Kira Astakhova. 2015. "Novel Signal-Enhancing Approaches for Optical Detection of Nucleic Acids—Going beyond Target Amplification" Chemosensors 3, no. 3: 224-240. https://doi.org/10.3390/chemosensors3030224