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Objective To evaluate whether virtual non-enhanced (VNE) images generated from the contrast-enhanced spectral CT images could replace true non-enhanced images (TNE) in pediatric abdominal imaging. Medthods A total of 35 children (26 boys and 9 girls, mean age: 9.3 years ± 3.17) underwent non-enhanced and enhanced abdominal CT using a 64-row CT scanner (GE Discovery CT 750 HD). The arterial phase (AP) of the enhanced scan was acquired using spectral CT with a low tube current (260mA). The non-enhanced and portal venous phase (PVP) images were acquired using conventional multi-slice spiral CT scans. The CT attenuation values, image noise and contrast to noise ratios (CNR) of the liver, kidney, pancreas, spleen and muscle were measured in VNE and TNE images. Two radiologists performed independent and blinded assessment of image quality. Effective radiation doses were calculated. Results There was no difference in CNR between VNE and TNE for liver (0.76±0.64 vs. 0.72±0.92), spleen (0.03±0.40 vs.0.02±0.40), kidney (1.21±0.41 vs. 1.27±0.34) and pancreas (0.09±0.48 vs.0.14±0.49) (all p>0.05). Mean CT attenuation values from VNE (at 70keV) were systematically lower than those on TNE (at 65keV) for all tissue types due to photon energy difference, but with good one-to-one correlation (liver: 63.99±6.39 vs. 70.31±8.71HU, spleen: 53.28±4.60 vs. 61.13±4.06HU, kidney: 38.04±4.05 vs. 45.27±3.81HU, pancreas: 51.67±4.52 vs. 58.32±5.10HU, and muscle: 55.31±3.66 vs. 61.59±3.42HU). Image noise was similar on VNE and TNE images (13.03±1.9 vs. 12.64±0.89HU). Image quality score for VNE was lower than TNE (4.17±0.09 vs. 4.72±0.07), but was acceptable for diagnosis. Effective radiation doses of the two-phase protocol (arterial phase with spectral CT imaging mode and portal-venous phase performed by multi-slice spiral CT); omitting the TNE scan was similar to the conventional three phase protocol (4.20±0.75 vs. 4.29±1.63mSv). Conclusion Image quality of VNE images is acceptable for diagnosis and CT numbers of VNE images correlate well with TNE images. TNE can be omitted from abdominal CT imaging in children, providing a simpler protocol.