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Method for detecting the presence of a target nucleic acid sequence in a sample

  • US 8,859,204 B2
  • Filed: 08/13/2007
  • Issued: 10/14/2014
  • Est. Priority Date: 04/17/1997
  • Status: Expired due to Fees
First Claim
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1. A method for detecting whether a target nucleic acid sequence is present in a biological sample, said method comprising:

  • loading sample portions of at least a first sample comprising nucleic acid into a plurality of respective sample chambers of a fluidic device, wherein said loaded sample portions have respective volumes ranging from 1 picoliter to 1 microliter, said sample chambers comprising means for minimizing diffusion of said sample portions such that when at least one sample portion loaded into a respective sample chamber contains a single molecule of a target nucleic acid sequence configured to produce amplified product of the target nucleic acid sequence in response to a primer-based amplification reaction, said at least one sample portion will attain a detectable concentration of amplified product of said target nucleic acid sequence within a portion of said respective sample chamber after being subjected to a primer-based amplification reaction with reagents configured and sufficient to perform primer-based amplification and fluorescence-based detection of said target nucleic acid sequence;

    simultaneously subjecting said sample portions in said sample chambers to the primer-based amplification reaction with said reagents sufficient to perform primer-based amplification and fluorescence-based detection of amplified product of said target nucleic acid sequence when present; and

    while in said sample chambers, detecting whether any of said sample portions contain amplified product of the target nucleic acid sequence after said subjecting.

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