The objective of our study was the determination of the influence of the sequential and spiral acquisition modes on the concordance and deviation of the calcium score on 64-slice multi-detector computed tomography (MDCT) scanners in comparison to electron beam tomography (EBT) as the gold standard. Our methods and materials were an anthropomorphic cardio CT phantom with different calcium inserts scanned in sequential and spiral acquisition modes on three identical 64-slice MDCT scanners of manufacturer A and on three identical 64-slice MDCT scanners of manufacturer B and on an EBT system. Every scan was repeated 30 times with and 15 times without a small random variation in the phantom position for both sequential and spiral modes. Significant differences were observed between EBT and 64-slice MDCT data for all inserts, both acquisition modes, and both manufacturers of MDCT systems. High regression coefficients (0.90-0.98) were found between the EBT and 64-slice MDCT data for both scoring methods and both systems with high correlation coefficients (R2>0.94). System A showed more significant differences between spiral and sequential mode than system B. Almost no differences were observed in scanners of the same manufacturer for the Agatston score and no differences for the Volume score. The deviations of the Agatston and Volume scores showed regression dependencies approximately equal to the square root of the absolute score. The Agatston and Volume scores obtained with 64-slice MDCT imaging are highly correlated with EBT-obtained scores but are significantly underestimated (-10% to -2%) for both sequential and spiral acquisition modes. System B is more independent of acquisition mode to calcium score than system A. The Volume score shows no intramanufacturer dependency and its use is advocated versus the Agatston score. Using the same cut points for MDCT-based calcium scores as for EBT-based calcium scores can result in classifying individuals into a too low risk category. System information and scanprotocol is therefore needed for every calcium score procedure to ensure a correct clinical interpretation of the obtained calcium score results.