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Flow cytometric detection method for DNA samples

Patent ·
OSTI ID:1019570
 [1];  [1];  [2]
  1. Livermore, CA
  2. Round Rock, TX
+ Show Author Affiliations
Disclosed herein are two methods for rapid multiplex analysis to determine the presence and identity of target DNA sequences within a DNA sample. Both methods use reporting DNA sequences, e.g., modified conventional Taqman.RTM. probes, to combine multiplex PCR amplification with microsphere-based hybridization using flow cytometry means of detection. Real-time PCR detection can also be incorporated. The first method uses a cyanine dye, such as, Cy3.TM., as the reporter linked to the 5' end of a reporting DNA sequence. The second method positions a reporter dye, e.g., FAM.TM. on the 3' end of the reporting DNA sequence and a quencher dye, e.g., TAMRA.TM., on the 5' end.
Research Organization:
Lawrence Livermore National Laboratory (LLNL), Livermore, CA (United States)
Sponsoring Organization:
USDOE
DOE Contract Number:
W-7405-ENG-48
Assignee:
Lawrence Livermore National Security, LLC (Livermore, CA)
Patent Number(s):
7,972,818
Application Number:
US Patent Application 11/454,478
OSTI ID:
1019570
Country of Publication:
United States
Language:
English

References (5)

A Reusable Flow-Through Polymerase Chain Reaction Instrument for the Continuous Monitoring of Infectious Biological Agents journal July 2003
Autonomous Detection of Aerosolized Biological Agents by Multiplexed Immunoassay with Polymerase Chain Reaction Confirmation journal January 2005
A Bead-Based Method for Multiplexed Identification and Quantitation of DNA Sequences Using Flow Cytometry journal October 2000
A multiplexed PCR-coupled liquid bead array for the simultaneous detection of four biothreat agents journal April 2005
Detection of specific polymerase chain reaction product by utilizing the 5'----3' exonuclease activity of Thermus aquaticus DNA polymerase. journal August 1991

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