[Objectives]  The infection of Nosema ceranae caused nosemiosis, but the mechanism of infection was unknown. Based on the previous work, this study was intended to further verify the expression level and sequencing results of nce-miR-15325, and to detect the relative expression levels of nce-miR-15325 and its target genes during the infection of Apis mellifera ligustica by Nosema ceranae. This study provided the basis for further exploring the function and mechanism of nce-miR-15325 in regulating infection [Methods]  Stem-loop RT-PCR and Sanger sequencing were employed to confirm the expression of, and to sequence, nce-miR-15325. Related software was used to predict and analyze relevant target genes. The expression profiles of nce-miR-15325 and its target genes were determined using RT-qPCR. [Results]  nce-miR-15325 was present and expressed in Nosema ceranae spores. Eleven target genes were predicted, of these, three, four, seven and four, respectively, were annotated to the KEGG, GO, Nr, and Swissprot databases. Compared to its expression at 1 dpi (1 day post-inoculation), the expression of nce-miR-23928 was unchanged at 2 dpi, but was progressively less at 4, 6, and 8 dpi (P<0.05). Compared to its expression at 1 dpi, the expression of the target gene NCCS was also progressively less (P<0.05) at 2, 4, 6 and 8 dpi, as was that of the target gene DTIII. [Conclusion]  These results confirm the existence and expression of nce-miR-15325 in Nosema ceranae spores, reveal its expression dynamics and that of its target genes during Nosema ceranae infection, and suggest that nce-miR-15325 potentially regulates the infection process via positive modulation of the expression of NCSS and DTIII.