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Comparative evaluation of HTG and TempO Seq targeted transcriptome profiling methods
Antonio Fernández-Serra1,2, Raquel López-Reig1,2, Ignacio Romero3
1Laboratory of Molecular Biology, Valencian Institute of Oncology, Valencia, Spain.
Scientific Reports
|January 24, 2026
Summary
TempO-Seq offers a viable alternative for targeted RNA sequencing in molecular oncology research. It shows comparable performance to the discontinued HTG EdgeSeq HTP platform for multi-gene biomarkers in FFPE samples.
Area of Science:
- Molecular biology
- Translational oncology
- Molecular oncology
Background:
- Transcriptomic profiling is crucial for molecular oncology, requiring reliable gene expression measurements.
- Targeted RNA sequencing offers advantages for formalin-fixed paraffin-embedded (FFPE) samples.
- The discontinuation of the HTG EdgeSeq HTP platform created a need for alternative targeted transcriptome analysis methods for FFPE specimens.
Purpose of the Study:
- To systematically compare the HTG EdgeSeq HTP platform with the TempO-Seq platform.
- To assess TempO-Seq's suitability as an alternative for targeted transcriptome analysis in molecular oncology research using FFPE samples.
Main Methods:
- Analysis of 21 FFPE endometrial cancer samples and three RNA reference controls.
- Evaluation of concordance between platforms across individual transcripts and multi-gene signatures.
- Systematic technical comparison of HTG EdgeSeq HTP and TempO-Seq platforms.
Main Results:
- Single-gene measurements showed limited concordance between the two platforms.
- Multi-gene biomarkers demonstrated substantially higher agreement between platforms.
- Aggregation across multiple probes mitigated platform-specific effects on biomarker agreement.
Conclusions:
- TempO-Seq provides comparable performance to HTG EdgeSeq HTP for multi-gene biomarkers.
- TempO-Seq is a robust alternative for targeted transcriptomic profiling of FFPE samples in molecular oncology research.
- The TempO-Seq platform can fill the technical gap left by the discontinuation of HTG EdgeSeq HTP.
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