In paternity testing, short tandem repeats (STRs) allele mismatches are often detected. Nowadays, polymerase chain reaction‐ and capillary electrophoresis (CE)‐based STR genotyping is the most commonly used method to distinguish alleles based on their length. However, it could not detect alleles of the same size with sequence differences. Massively parallel sequencing (MPS) can determine not only allele sizes but also sequences, which could explain the causes of allele mismatches. Additionally, more types of genetic markers can be detected in a single assay, which increases the discriminatory power and facilitates the analysis of paternity tests. In this study, we analyzed 11 cases with homozygous allele mismatches from routine DNA trio paternity tests using the CE platform. Samples were sequenced using the ForenSeq DNA Signature Prep Kit and the MiSeq FGx Sequencing System. The results show that of the eight father–child mismatch cases and three mother–child mismatch cases, five cases with D5S818 and D8S1179 and one case at D13S317 were classified as non‐amplification. The other three cases and two cases could be defined as mutations. This study suggests that MPS‐based STR genotyping can provide additional information that allows more accurate interpretation of allelic mismatches in paternity testing.