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The two main screening tests during pregnancy are those for chromosomal abnormalities and neural tube defects(NTDs). In particular, for NTDs, measurement of maternal serum alpha-fetoprotein (MSAFP) levels early in the secondtrimester (15-18 weeks of gestation) has been considered the gold standard screening test for the past 4 decades. However, with remarkable technological advancements and the widespread use of ultrasound during those periods,mid-trimester ultrasonography has gradually replaced the role of measuring MSAFP levels as a screening method forNTDs. This change was initiated more about 10 years ago in some countries, which have issued national guidelinesto use mid-trimester ultrasonography instead of measuring MSAFP levels as a prenatal screening method for NTDs. However, no significant changes have occurred in Korea, where second-trimester ultrasonography is routinelyperformed with high-quality equipment. We aimed to provide information regarding the importance of changing thescreening method for NTDs from MSAFP measurement to ultrasonography, and to detail methods of implementingmid-trimester ultrasonography for screening purposes. We also share our experience of operating a prenataldiagnostic program for NTDs without using MSAFP for more than 15 years.