Good hand hygiene has proven to be essential in reducing the uncontrolled spread of human pathogens. Cold atmospheric plasma (CAP) may provide an alternative to disinfecting hands with ethanol-based handrubs when handwashing facilities are unavailable. CAP can be safely applied to the skin if the energy is well controlled. In this study, radio frequency (RF) and direct current (DC) plasma sources were built with a pin-to-mesh electrodes configuration inside a fused silica tube with a 5 mm inner diameter. Microbiological assays based on EN 13697:2015+A1:2019 using Escherichia coli DSM 682 or Staphylococcus epidermidis DSM 20044 were used to examine the antimicrobial effect of various plasma conditions. Metal and silicone disks that model skin were used as inoculation matrices. The prototype air RF CAP achieved significant disinfection in the MHz range on stainless steel and silicone substrates. This is equivalent to half the performance of direct current CAP, which is only effective on conductive substrates. Using only electricity and air CAP could, with further optimization to increase its efficacy, replace or complement current hand disinfection methods, and mitigate the economic burden of public health crises in the future.