Qualitative and quantitative detection of microbial nucleic acids

Qualitative and quantitative detection of microbial nucleic acids

  • CN 103,069,007 A
  • Filed: 07/27/2011
  • Published: 04/24/2013
  • Est. Priority Date: 07/29/2010
  • Status: Active Application
First Claim
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1. method that is used for detecting and quantizing simultaneously the microbial nucleic acids of biological sample, described method comprises:

  • A) separate the also described microbial nucleic acids of purifying,B) provide reaction mixture, its comprise at least the first contrast nucleic acid, with the unique sequences of described microbial nucleic acids part and with one or more primer pairs of the unique sequences part specific hybrid of described contrast nucleic acid and with each probe by sequence-specific hybridization of described one or more primer pairs amplifications, wherein said microbial nucleic acids and described contrast nucleic acid and different probe hybridizationC) add described biological sample to described reaction mixture,D) implement one or more circulation steps, wherein circulation step comprises amplification step, if described amplification step comprises described microbial nucleic acids and is present in the described sample, then generate one or more from the derivative amplified production of described microbial nucleic acids, and generate from the derivative amplified production of described contrast nucleic acid, and wherein circulation step comprises hybridization step, described hybridization step comprises sequence and the described probe hybridization that makes by described primer pair amplification, wherein said probe is with donor fluorescence module and accept accordingly the fluorescence module marks, and every kind of probe carries different fluorescence dyesE) detect and measure by described amplified production generate and to the proportional fluorescent signal of the concentration of described contrast nucleic acid and described microbial nucleic acids, even the existence of the amplified production of wherein said contrast nucleic acid is increased in the described reaction mixture of situation indicating of the amplified production that lacks described microbial nucleic acids.

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