Method for quantitative end-point PCR
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United States of America Patent
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Issued Date -
Jan 20, 2005
app pub date -
Oct 17, 2003
filing date -
Oct 18, 2002
priority date (Note) -
Abandoned
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Abstract
The present invention provides a sensitive and robust analytical approach to identifying and quantifying multiple pathogens within a single complex environmental sample. Numerous nucleic acid signatures may be screened and quantified within a single reaction tube using polymerase chain reaction (PCR) and chromatographically analyzing the amplification products with microchannel fluidic (e.g. Agilent 2100 Bioanalyzer, or Caliper AMS-90) or reverse-phase ion-pairing high-performance liquid chromatography RP IP HPLC (e.g. Transgenomic WAVE) instruments. The method may be employed in a multiplex fashion to allow identification and quantification of multiple combinations of up to five different nucleic acid signatures simultaneously within a single multiplex PCR reaction tube. This approach is quantitative across a dynamic range of up to five orders of magnitude. This method is suitable for target nucleic acid analysis in medium or high-throughput contexts such as routine clinical diagnotics or environmental monitoring. The method is also suitable for pathogen monitoring or surveillance in a biodefense context.
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