All-natural rubber is harvested from rubber trees (Hevea brasiliensis Muell. Arg.) by traditional tapping knives, so rubber tapping still heavily relies on labor. Therefore, this study explored a novel, hand-held mechanical rubber tapping machine for rubber tree harvesting. In this study, a mechanical tapping cutter with a vertical blade and adjustable guide was first described. The response surface method was applied to evaluate factors affecting the tapping effect. The experimental values were in close agreement with the predicted value. Machine-tapped latex was comparable in quality to hand-tapped latex. Based on the single-factor results, the response surface method (RSM) and the center combined rotation design (CCRD) optimization method were adopted to explore the influence of three factors influencing vertical blade height (A), cutting force (B), and spiral angle (C) on the tapping effect. Regarding the cutting rate of the old rubber line (Y1), cutting time (Y2), latex flow rate (Y3), and average cutting current (Y4) as evaluation indexes of the tapping effect, an optimization scheme was determined. The quadratic model fits for all the responses. The test results showed that the main factors affecting Y1, Y2, Y3, and Y4 were A and B, B, A and C, and B, respectively. Under optimal conditions, the influencing factors of A, B, and C were 10.24 mm, 51.67 N, and 24.77°, respectively, when the evaluation index values of Y1, Y2, Y3, and Y4 were 98%, 8.65 mL/5 min, 9.00 s, and 1.16 A. The range of the relative error between the experimental and predicted results was from −11.11% to 11.11%. According to the optimized treatment scheme, a comparison test was designed between mechanical and manual rubber tapping tools. To verify the availability and effect of the mechanical tapping method preliminarily, the important rubber tapping evaluation indexes included bark thickness, bark excision, latex flow time, cutting time, ash content, and cutting depth, which were selected to serve as a comparison test. There was no significant difference between hand and mechanical methods, except ash content (p < 0.05) and cutting time (p < 0.01). The mechanical tapping machine proposed in this study is meaningful to improve cutting efficiency, practicality, and operability. Furthermore, it provides crucial theoretical references for the development of intelligent tapping machines.