Background In Dutch hospitals a 6-point questionnaire is mandatory for risk-assessment to identify carriers of multidrug-resistant organisms (MDRO) at the time of hospitalization. Presence of one or more risk factors is followed by microbiological culturing and pre-emptive isolation. We evaluated the test characteristics of this screening tool in identifying new MDRO carriers. MethodsA cross-sectional study using routinely collected healthcare data was performed in a Dutch tertiary hospital between 1 January 2015 and 1 August 2019 including all admissions with an MDRO risk assessment performed on the day of admission. MDRO risk-assessment included: (1) known MDRO carriage, (2) previous hospitalization in another Dutch hospital during a known outbreak, (3) previous hospitalization in a foreign hospital, (4) living in an asylum centre, (5) professional exposure to livestock farming and (6) household membership of a methicillin-resistant Staphylococcus aureus (MRSA) carrier. Sensitivity of the risk assessment was estimated by comparing observed prevalence of newly detected MDRO carriage to expected prevalence of carriage in the Dutch population upon hospital admission. Results 144,051 hospital admissions of 84,485 unique patients were included. In total, 4,480 (3.1%) admissions had a positive MDRO risk-assessment (i.e. ≥1 risk factors present). In 1,516 (34%) admissions microbiological screening was performed, of which 341 (23%) yielded MDRO. 81 patients were categorized as new MDRO carriers, as identified through MDRO risk-assessment, reflecting 0.06% (95% CI: 0.04%–0.07%) of all admissions and 1.8% (95% CI: 1.4%–2.2%) of those with positive risk assessment. MDRO included ESBL-producing and/or multidrug-resistant Enterobacterales (n=52, 64%), MRSA (n=26, 32%), carbapenem-resistant Enterobacterales (CRE) (n=2, 3%) and VRE (n=1, 1%). The numbers of “MDRO risk-assessments needed to perform” and individual “MDRO risk-assessment questions needed to ask” to detect one new MDRO carrier upon admission were 1,778 and 10,420, respectively. Estimated sensitivities of the risk-assessment for detecting MDRO carriage were <1%, for ESBL-E and VRE, <2% for CRE and 18% for MRSA. Conclusions The number of risk-assessments needed to perform to detect one new MDRO carrier upon hospital admission was high, and the vast majority of carriers most likely remained undetected. The current MDRO risk assessment upon admission strategy needs thorough reconsideration.