Congenital heart disease (CHD) is the most common developmental anomaly, affecting approximately 1% of all newborns. Genetic factors play an important role in CHD's development. Germline mutations in NK2 transcription factor related, locus 5 (NKX2.5) have been identified as the factors responsible for various forms of CHD. In this study, we investigated mutations of the NKX2.5 gene's coding region in 230 nonsyndromic CHD patients belonging to the Chinese Han nationality by PCR, denaturing high-performance liquid chromatography, and sequencing. Pathogenic mutations were not found among the patients. Two known single-nucleotide polymorphisms (rs2277923 and rs3729753) were detected, but the differences in the allele and genotype frequencies were insignificant between CHD and the controls (p > 0.05). The data we gathered suggest that NKX2.5 mutations are highly rare in CHD patients of the Chinese Han nationality. Therefore, NKX2.5 mutation investigation should be limited within a number of familial and special phenotype of CHD in Chinese patients.