Method for detecting nucleic acid sequences using competitive am

Details Method for detecting nucleic acid sequences using competitive am Method for detecting nucleic acid sequences using competitive am

1995-06-07

1997-09-16

Zitomer, Stephanie W.

Chemistry: molecular biology and microbiology

Measuring or testing process involving enzymes or...

Involving nucleic acid

536 243, 435 912, C12Q 168, C12P 1934, C07H 2104

Patent

active

056679742

ABSTRACT:
The present invention provides a method for quantitatively detecting the amount of a target nucleic acid sequence which may be present in a test sample. A test sample which may contain a target nucleic acid sequence comprising target sequences X and Y is contacted with two primer sets, the first set being specific for target X and the second set being specific for target Y. The test sample also is contacted at the same time with an internal standard sequence IS, which is substantially derived from a combination of the first and second target sequences, and its corresponding oligonucleotide primers. Haptens are associated with the oligonucleotide primer sets in such a way that amplified target sequence products X and Y are detected by capture on a solid phase to which anti-hapten capture reagents are attached. A signal ratio of (X+Y)/S is determined to quantitate the amount of the target nucleic acid sequence contained in the sample.

REFERENCES:
patent: 5219727 (1993-06-01), Wang et al.
patent: 5427930 (1995-06-01), Birkenmeyer et al.
P. Fang et al: "Simultaneous Analysis of Mutant and Normal Alleles for Multiple Cystic Fibrosis Mutations by the Ligase Chain Reaction." Human Mutation 6:144-157 (1995).
B. Goswami et al., "Competitor Template RNA For Detection and Quantitation of Hepatitis A Virus by PCR," Biotechniques 16(1): 114-121 (1994).

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