Objective To evaluate the effect of active screening and intervention of multidrug-resistant organisms (MDROs) on control nosocomial infection in the general intensive care unit (ICU). Methods A non-concurrent control trial was conducted in patients hospitalized in the ICU for more than 24 hours in the Second Affiliated Hospital of Fujian Medical University. Patients underwent active screening of MDROs for nasal vestibular swab, throat swab and rectal swab combined with further intensive intervention for patients with positive screening result during Sept. 2014 to Aug. 2015 were included as an intervention group, patients only underwent active screening during Sept. 2013 to Aug. 2014 were included as a screening group, and patients without undergoing active screening during Sept. 2012 to Aug. 2013 were as a control group. SPSS 19.0 software was used to compare the hospital infection rate and the infection rate of MDROs among the three groups. Results A total of 1 773 patients were included, of which 655 patients were in the intervention group, 515 patients were in the screening group, and 603 patients were in the control group. The difference of hospital infection rates among the three groups was statistically significant (χ2=21.087, P < 0.001), and further pairwise comparison results showed that the intervention group was lower than the screening group (χ2=5.891, P=0.015), and the screening group was lower than the control group (χ2=4.259, P=0.039). The adjustment daily infection rate of the intervention group, screening group and control group were 6.69‰, 10.88‰, and 15.39‰, respectively. The difference of MDROs hospital infection rates among the three groups was statistically significant (χ2=21.039, P < 0.001), and further pairwise comparison results showed that the intervention group was lower than the screening group (χ2=5.936, P=0.015), and the screening group was lower than the control group (χ2=5.798, P=0.016). The MDROs thousand daily infection rate of the intervention group was lower than that of the screening group (3.90‰ vs. 7.30‰, χ2=5.999, P=0.014). Conclusion The active screening plus intensive intervention of MDROs can effectively reduce the incidence rates of nosocomial infections and MDROs infections in ICU.