This prospective study assessed the clinical application of a nonfasting low-density lipoprotein cholesterol measure in a pediatric primary care population. A homogenous direct low-density lipoprotein cholesterol assay was tested in healthy children, aged 4 to 12 years, at risk for hyperlipidemia, as defined by American Academy of Pediatrics, and including patients with incomplete family histories. This nonfasting homogeneous assay was comparable to modified beta quantification, the gold standard reference method of measuring low-density lipoprotein cholesterol, and the current recommended screening method, total cholesterol. Results from the study suggest that this nonfasting assay can provide more direct low-density lipoprotein cholesterol measurements for healthy children, with improved clinical utility and greater overall patient convenience in testing for important cholesterol abnormalities.