Abstract
Background: Recently, various sets of protein biomarkers have been discovered in important diseases such as cancers, brain stroke and heart attack. However, clinical validation is difficult and time-consuming by individual assays or because of very low concentrations at early stages of the diseases. We have developed assay technology through an innovative modification of the immuno-PCR method for the super-sensitive and multiplex detection of target biomarkers.
Methods: In the assay technology, each different oligo-tag simultaneously detects multiplex protein targets with extremely high-level sensitivity in a dose-dependent manner by qRT-PCR (maximum: three plexes). In this study, we measured specific secreted protein concentrations in the culture supernatant of a 24-h culture of transfected SH-SY5Y cells with MUSTag.
Results: There was a significant increase in the protein level of tumor necrosis factor (TNF)-α measured with extremely high-level sensitivity (≥10 pg/mL). Compared with negative controls, the levels of TNF-α increased from 16.9 to 28.1 pg/mL (p = 0.011).
Conclusion: We suggest that our assay technology might be of clinical value in treating patients with cancer, cerebral ischemia, or patients who need a prompt and predictive diagnosis for adequate treatment.
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Acknowledgments
We are grateful to A. Sakurai, T. Hashimoto, and A. Endler for suggestions and critical reading of the manuscript; N. Makisaka for technical assistance; and all the members of our laboratory for their input during the course of this work.
Conflict of InterestWe declare that we have no conflict of interest.
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Miyashita, R. et al. (2013). Cytokine Marker Measurement in Human Neuroblastoma Cells with Supersensitive and Multiplex Assay: MUSTag Technology. In: Katayama, Y., Maeda, T., Kuroiwa, T. (eds) Brain Edema XV. Acta Neurochirurgica Supplement, vol 118. Springer, Vienna. https://doi.org/10.1007/978-3-7091-1434-6_62
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DOI: https://doi.org/10.1007/978-3-7091-1434-6_62
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