Aim To investigate the effect on implementation of standard procedure of recombinant tissue-type plasminogen activator (rt-PA) intravenous thrombolysis in patients with acute ischemic stroke (AIS). Methods One hundred and sixty-four AIS patients were enrolled in this study. Thirty-four cases were assigned into the before-implementation group and one hundred and thirty into the after-implementation group. Door-to-needle time (DNT), NIHSS scores and modified Rankin scale (mRS) scores of the patients in the two groups were analyzed. The durations of every step in the treatment procedure were statistically compared between the two groups. Results The average DNT in the after-implementation group [(81.31±15.13) min] was significantly shorter than that in the before-implementation group [(111.76±24.84) min] (P＜0.001). The discharge NIHSS scores declined remarkably compared with baseline NIHSS scores in both two groups, and the difference had statistical significance (P＜0.01). The discharge NIHSS scores and mRS scores in the after-implementation group was significantly lower than that in the before-implementation group (P＜0.01). The time interval between arrival at emergent department and beginning CT scan in the two groups was not statistically significant. The time interval between arrival at emergent department and starting the procedure by telephone, consulting chief resident physician and signing the informed consents were significantly shorter in the after-implementation group (P＜0.01). Conclusion Implementing MDT model of rt-PA intravenous thrombolysis standarding procedure contributes to shortening the DNT and improving the therapeutic effects.