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Wide dynamic range nucleic acid detection using an aggregate primer series

  • US 5,858,732 A
  • Filed: 08/23/1996
  • Issued: 01/12/1999
  • Est. Priority Date: 05/19/1995
  • Status: Expired due to Fees
First Claim
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1. A method for determining an approximate amount of a target nucleic acid sequence in a test sample comprising the steps of:

  • (a) contacting said test sample with a nucleic acid amplification reaction mixture comprising a first primer set capable of hybridizing with a first sub-target region of said target nucleic acid sequence, and a second primer set capable of hybridizing with a second sub-target region of said target nucleic acid sequence, wherein said first and said second sub-target regions are different and wherein said first and said second primer sets are selected such that(i) said first primer set is capable of producing a detectable amplicon at or above a first pre-selected threshold concentration of said target nucleic acid sequence,(ii) said second primer set is capable of producing a detectable amplicon at or above a second pre-selected threshold concentration of said target nucleic acid sequence, and(iii) said first and second threshold concentrations are different from one another;

    (b) subjecting the reaction mixture to amplification conditions sufficient to produce a detectable amplicon from said primer sets when said test sample contains a concentration of target nucleic acid sequence which is at or above the different threshold concentrations at which the primer sets are capable of producing a detectable amplicon; and

    (c) detecting whether amplicons are produced in said reaction mixture from said primer sets so as to determine whether the test sample contains a concentration of target nucleic acid sequence at or above said threshold concentrations corresponding to said primer sets.

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