Background and objective: The disadvantage of the traditional 20-m multistage shuttle run test (MST) is that it requires a long space for measurements and does not include various age groups to develop the test. Therefore, we developed a new MST to improve the spatial limitation by reducing the measurement to a 10-m distance and to resolve the bias via uniform distributions of gender and age. Material and methods: Study subjects included 120 healthy adults (60 males and 60 females) aged 20 to 50 years. All subjects performed a graded maximal exercise test (GXT) and a 10-m MST at five-day intervals. We developed a regression model using 70% of the subject’s data and performed a cross-validation test using 30% of the data. Results: The male regression model’s coefficient of determination (R{}^2) was 58.8%, and the standard error of estimation (SEE) was 4.17 mL/kg/min. The female regression model’s R{}^2 was 69.2%, and the SEE was 3.39 mL/kg/min. The 10-m MST showed a high correlation with GXT on the VO{}_2max (males: 0.816; females: 0.821). In the cross-validation test for the developed regression models, the male’s SEE was 4.38 mL/kg/min, and the female’s SEE was 4.56 mL/kg/min. Conclusion: Thus, the 10-m MST is an accurate and valid method for estimating the VO{}_2max. Therefore, the 10-m MST developed by us can be used when the existing 20-m MST cannot be used due to spatial limitations and can be applied to both men and women in their 20s and 50s.