Hydrogel-compartmentalized heterogeneous amplification for viral digital genotyping

Accurate viral genotyping is significant for exploring clinical manifestations and treatment options. The classical nucleic acid amplification within homogeneous system merely reflects the average characteristics of all molecules in samples, which prevents the simultaneous analysis for sample with viruses of different genotypes. Herein, we propose hydrogel-compartmentalized nucleic acid amplification and introduce restriction enzyme to establish a competitive amplification reaction. This system localizes high molecular-weight viral nucleic acids and allows single-molecule heterogeneous amplification in response to various endonuclease digestion. With this strategy, we could provide accurate information for digital genotyping of Hepatitis B virus. Considering its well-performance and easy-to-use, the hydrogel-compartmentalized heterogeneous amplification system should be a promising tool for digital genotyping, and will be of great help to other genetic analysis.