TABLE 3.
Advantages and disadvantages of various point mutation detection techniques.
| Based point mutations detection | Advantages | Disadvantages |
|---|---|---|
| Sanger | Technologically mature, High accuracy | Low sensitivity, High cost |
| NGS | High-throughput, High sensitivity, Well-suited for complex mutations | Complex data analysis, High error rate, Requires relatively specialized equipment |
| TGS | Long-reads, More suitable for detecting large-scale variations | High cost, Low adoption rate, Time-consuming |
| Guided enzymes | Technological maturity, Programmability, Low cost, Broad application potential | Limitations of PAM sequences, Off-target effects, Delivery challenges, Immune response |
| Ligase | Low sample requirement, Multiplex detection capability, High sensitivity | Thermal instability, Low catalytic efficiency, Complex probe design |
| DNAzyme | Low cost, Programmability, High stability, Low immunogenicity | Low catalytic efficiency, Metal ion-dependent, Complex synthesis |
| Nanozyme | High stability, Versatile design, Easy to synthesize, High sensitivity | Potential toxicity, Challenges in large-scale production, Low specificity |
| Enzyme-free | Low cost, Programmability, Easy to operate | High design complexity, Low sensitivity, Restricted in vivo application |
| Colorimetry | Simple and rapid operation, Visual readout, High sensitivity | Background interference, Sensitive to reaction conditions, Difficult to quantify |
| Fluorescence methods | High sensitivity, Quantitative analysis capability, Multiplex detection | Background interference, High cost, Requires sophisticated equipment |
| Electrochemical methods | High sensitivity and specificity, Rapid detection | Complex operational procedure, Limited clinical applicability |
| SERS | Multiplex detection, High sensitivity and specificity, Low sample consumption, Rich molecular information | Requires sophisticated equipment, Difficult to quantitative detection, Complex signal analysis, Requires sophisticated probe design |
| PNA | High affinity, Resistant to enzymatic degradation, High specificity | Poor aqueous solubility, Inefficient cellular internalization, High production costs |
| LNA | Enhanced binding affinity, High stability, Compatibility with various detection platforms | Probe structural rigidity, Potential toxicity risks, Complex probe design |
| Pure nucleic acid probe | Programmable structure and function, Multiplatform compatibility | Insufficient dynamic regulation, Limited interference resistance |
| AI-assisted | High accuracy, High efficiency, Minimal errors and biases | Technologically immature, Black box effect, High technical requirements |