Contact-killing surfaces with the ability to rapidly adsorb and kill microorganisms are desperately needed since the rapid outbreak of multidrug-resistant (MDR) bacteria poses a serious threat to human health. Therefore, a series of amphiphilic nanoengineered polyquaterniums (ANPQs) were synthesized, and immobilizing ANPQs onto equipment surfaces provided a simple method for preventing microbial infections. The strong charge-positive property of ANPQ offered the possibility of rapid adsorption and efficient killing, such that all bacteria are adsorbed after 10 seconds of contact with ANPQ-treated fabrics, and more than 99.99% of pathogens are killed within 30 seconds. Surprisingly, the adsorption-killing mechanism made it difficult for bacteria to develop resistance to ANPQ coating, even after long-term repeated treatment. Importantly, in a Methicillin-resistant Staphylococcus aureus infection model, ANPQ-treated fabrics exhibited a potent anti-infectious performance while remaining nontoxic. It is envisaged that the strategy of using ANPQ coating undoubtedly provides a promising candidate for fighting MDR strains.