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Flow cytometric detection of specific DNA sequences in human lymphocytes

  The cells that harbor specific DNA sequences of interest cannot be detected by current flow cytometry technologies. Since DNA is present at one or two copies per cell, target cells that harbor DNA labeled with FISH probe are hardly detected by flow cytometry. Although many signal amplification technologies have been developed, the flow cytometry assay that can detect cells harboring specific DNA sequences has not been established. We have overcome the limitation by target amplification rather than signal amplification and developed the novel flow cytometry assay that enables to detect cells harboring specific DNA sequences of interest. The sensitivity of the assay is high enough to detect single-copy DNA target in every cell. The method developed by us is a significant advance in the flow cytometry technology, because not only proteins but also DNA sequences of interest can be analyzed at the single cell level by flow cytometry. The method would be of broad interest because flow cytometry is generally used in research and clinical practice.

Naoki Uno, Norihito Kaku, Yoshitomo Morinaga, Hiroo Hasegawa, Katsunori Yanagihara
Flow cytometry assay for the detection of single-copy DNA in human lymphocytes
Nucleic Acids Research, gkaa515; 2020
https://doi.org/10.1093/nar/gkaa515