Genome-wide identification and characterization of a panel of house-keeping genes in Schistosoma japonicum

Quantitative real-time polymerase chain reaction (qPCR), as one of the most sensitive and precise gene expression analysis methods, is frequently used to validate data obtained in high-through-put assays. qPCR requires reference genes with stable transcription for accurate normalization. However, no systematic studies on such genes have been performed in the genus Schistosoma japonicum. In this study, eight novel candidate genes selected from a microarray analysis and four commonly used reference genes were systematically validated in a series of qPCR experiments. Based on the results of geNorm, Normfinder, BestKeeper, and the comparative delta-cycle threshold (ΔCT) integrated analysis, the genes PSMD4, NDUFV2, and TPC2L were found to be most stably expressed in all S. japonicum developmental stages; meanwhile, ACTB and TUBA were found as the least stably expressed genes. This study provided, at the first time, data for genes that can be explored as reliable references in transcriptomic analysis of S. japonicum.

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► Eight novel housekeeping genes in S. japonicum were identified and validated.
► The expression stability of the genes was evaluated by four algorithms.
► PSMD4, NDUFV2 and TPC2L were found to be most stably expressed in S. japonicum.
► PSMD4, NDUFV2 and TPC2L can be explored as reference genes in S. japonicum.