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- W4313365701 abstract "Abstract Immune repertoire amplification of T cell receptors (TCRs) coupled with high throughput sequencing provides detailed insight into the immune system. A common starting sample type is total RNA extracted from whole blood by venipuncture. However, the costs, participant burden, regulatory constraints, and logistics associated with venipuncture and RNA handling are major barriers to clinical application or community-based research on various diseases. This study aims to identify TCR repertoires from dry blood spots (DBS), a method that could help collect real-world data for biomarker applications. Finger-prick blood was collected onto a Whatman filter card, and RNA was extracted from DBS. Fully automated multiplex PCR was performed to generate a TCRβ chain library for next generation sequencing (NGS) analysis of unique CDR3s (uCDR3). According to statistical analysis and laboratory confirmation, forty 2-mm punch disks from the filter cards were enough to detect the shared top clones and have a strong correlation in uCDR3 discovery with whole blood. uCDR3 discovery was not affected by storage temperatures (room temperature or −20°C) or storage durations (1, 14, and 28 days) when compared to whole blood. About 74 – 90% of top 50 uCDR3 clones of whole blood could also be detected from DBS. The DBS-based TCR repertoire profiling method is minimally invasive, provides convenient sampling, and incorporates fully automated library preparation. The system is sensitive to low RNA input, and the results are highly correlated with whole blood uCDR3 discovery allowing study scale-up to better understand the relationship and mutual influences between the immune repertoire and various disease states." @default.
- W4313365701 created "2023-01-06" @default.
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- W4313365701 date "2019-05-01" @default.
- W4313365701 modified "2023-10-14" @default.
- W4313365701 title "T cell receptor repertoire detection from dry blood spots" @default.
- W4313365701 doi "https://doi.org/10.4049/jimmunol.202.supp.131.24" @default.
- W4313365701 hasPublicationYear "2019" @default.
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