Preservation of mRNA from Isolated Circulating Tumour Cells

It is well documented that TransFix treated blood is compatible with isolation of circulating tumour cells, but new work published by Baird et al. describes a novel method of isolating CTCs, and the first published method of extracting mRNA from TransFix-treated samples for RT-qPCR analysis.

  • Baird et al. (2019) Enumeration of rare cells in whole blood by signal ion emission reactive release amplification (SIERRA) with same-sample RNA analysis. Anal. Chem 91 (3); 2028-2034. https://doi.org/10.1021/acs.analchem.8b04446

The Authors present a platform capable of detecting less than 30 spiked circulating tumour cells from a whole blood sample collected in TransFix CTC-TVT tubes. Their technology is based on size-exclusion filtration, microfluidic sample handling, and mass spectrometric detection through signal ion emission reactive release amplification (SIERRA), and demonstrate compatibility with PCR-based gene assays.

The authors show that the isolated, TransFix-treated cells, could then be used for downstream gene analysis provided a “pre-treatment” step was carried out as described in the paper. This resulted in a mRNA yield 100x greater than untreated TransFixed samples. TransFix allowed the authors to store samples between 24-120 h at room temperature before analysis.

For more examples of publications referencing the use of TransFix, visit our bibliography section!

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TransFix Product Format

Circulating Tumour Cell TransFix/EDTA Vacuum Blood Collection Tubes (CTC-TVTs)Circulating Tumour Cell TransFix/EDTA Vacuum Blood Collection Tubes (CTC-TVTs) are 9ml evacuated tubes prefilled with TransFix. CTC-TVTs are specifically designed for the optimal collection and stabilisation of CTCs in whole human blood. They allow stabilisation of CTCs for up to 5 days immediately preventing the degradation of CTCs and CTC antigens, allowing time to isolate and analyse these rare cells.

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Please note that the published applications in this blog do not relate to an IVD product claim. Cytomark have not verified these findings.

 

 

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