Abstract The effects of finite β (the ratio of plasma kinetic pressure to magnetic pressure) and three-dimensional (3D) magnetic perturbations (MPs) on the instability of toroidal ion temperature gradient (ITG) mode are studied in this work. The expression of ion magnetic drift frequency ω_di modified by the effects from both finite β and 3D MPs is firstly derived based on the local 3D equilibrium model. Then, under the assumptions of adiabatic electrons and localized mode structure around the outboard mid-plane (ϑ_p=0), the dispersion equation of the long wavelength toroidal ITG mode with considering the parallel ion dynamics is derived and solved. The results show that the distribution of ω_di around the outboard mid-plane, including both ω_(di,0)=├ ω_di ┤|_(ϑ_p=0) and ω_(di,0)^''≡├ (d^2 ω_di \/dϑ_p^2 )┤|_(ϑ_p=0) (twist parameter quantifying the degree of concave or convex of ω_di), is the key for affecting the toroidal ITG mode instability. The diamagnetic effects from finite β and the effects from 3D MPs can suppress the instability by reducing |ω_(di,0) |. Via reducing |ω_(di,0)^'' | under the prerequisite of unchanged sign of ω_(di,0)^'', there also exist stabilization effects on the instability from 3D MPs and the modification of local magnetic shear by finite β effects. In addition, the stabilization effects induced by reducing |ω_(di,0)^'' | are closely associated with the global magnetic shear. The mechanisms for the effects of finite β and 3D MPs on the instability of toroidal ITG mode revealed in this work are helpful to the comprehensive understanding of the relationship between internal kink mode induced non-axisymmetric flux surface distortion and internal transport barrier physics in tokamak plasmas.