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Twenty-nine opioid-exposed and 26 nonopioid-exposed neonates received neonatal abstinence syndrome (NAS) assessment by an examiner blinded to group status twice daily over the first two postnatal days. The opioid-exposed group had higher mean NAS scores than the nonopioid-exposed group. A 3-sign index, consisting of hyperactive moro reflex, mild tremors when undisturbed, and increased muscle tone, showed excellent discrimination between groups. The use of a 3-sign screening index in the days immediately after birth may provide a cost-effective mechanism for the identification of opioid-exposed infants, particularly in infants of women for whom identification of status as a substance user may not be immediately evident. Although a potentially useful screening tool, the 3-sign screening tool should not replace the full assessment of the opioid-exposed infant after birth.