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Bacterial infections and the emergence of drug-resistant bacteria threaten human health. Photothermal sterilization has the advantages of temporal and spatial control, no drug resistance, and high efficiency and speed. Photothermal therapy (PTT) causes cell death by means of heat generation. In this work, V2CTx nanosheets are successfully prepared by an improved etching method and the biocompatibility is verified by cytotoxicity. The experiments show that the photothermal properties of V2CTx materials have a significant positive correlation with laser power and material concentration. Only 20 μg/mL of V2CTx can be heated above 70 °C in just 5 min. Photothermal conversion efficiency of V2CTx reaches 45.15% and has good photothermal stability. Taking the typical Gram-negative Escherichia coli (E. coli) and Gram-positive Staphylococcus aureus (S. aureus) as experimental bacteria. The results prove that V2CTx itself is a good antibacterial agent, but antibacterial need more than 4 h of culture time. Instead, the photothermal sterilization method shows only 0.125% and 12.72% survival rates for S. aureus and E. coli within 8 min, respectively. Therefore, the prepare V2CTx in this paper only needs 20 μg/mL to achieve efficient photothermal sterilization, which provides a good prospect for clinical treatment of diseases caused by multidrug-resistant bacteria. [ABSTRACT FROM AUTHOR]