Patent ID: 9670540
Date: 2017-06-06
CPC Classifications: C12Q,G01N

Claim:
1. A method for identifying one or more of a plurality of target nucleotide sequences in a sample comprising: providing a sample potentially containing one or more target nucleotide sequences and/or complements thereof, each target nucleotide sequence comprising a first, second, and third target portion, wherein the second target portion is between the first and third target portions; providing one or more oligonucleotide primer sets, each set characterized by (a) a first oligonucleotide primer comprising a first primer portion complementary to the first target portion and a second primer portion that is the same as the second target portion, wherein the second primer portion is 5′ to the first primer portion, and (b) a second oligonucleotide primer comprising a third primer portion that is the same as the third target portion; providing a first polymerase; blending the sample, the oligonucleotide primer sets, and the first polymerase to form a polymerase extension reaction mixture; subjecting the polymerase extension reaction mixture to a hybridization treatment, wherein the first and third primer portions of the first and second oligonucleotide primers, respectively, hybridize to their respective complementary portions of the target nucleotide sequence or complement thereof, and an extension treatment, wherein the hybridized oligonucleotide primers extend to form primary primer extension products; denaturing the primary primer extension products from the target nucleotide sequences and complements thereof; providing a second polymerase; blending the polymerase extension reaction mixture after said subjecting it to the hybridization treatment and the extension treatment and the second polymerase to form a polymerase amplification reaction mixture; subjecting the polymerase amplification reaction mixture to one or more polymerase amplification reaction cycles comprising a denaturation treatment, wherein hybridized nucleic acid sequences are separated, a hybridization treatment, wherein the first and third primer portions of the first and second oligonucleotide primers, respectively, hybridize to their respective complementary portions of the primary primer extension products and an extension treatment, wherein hybridized oligonucleotide primers extend to form first oligonucleotide primer extension products and second oligonucleotide primer extension products; and subjecting the first and/or second oligonucleotide primer extension products to a sequencing reaction to identify the presence of one or more target nucleotide sequences in the sample.