Patent ID: 8192939
Filing Date: 2012-06-05
Classification: C12Q

Abstract:
1. A method for determining the presence of a viral target nucleic acid molecule in a sample containing a virus, the method comprising: (a) hybridizing a reverse primer to the viral target nucleic acid molecule under conditions suitable for carrying out a primer extension chain reaction; (b) extending the reverse primer using the viral target nucleic acid molecule as a template to form a reverse primer extension product, wherein the reverse primer joined to the reverse primer extension product constitutes a reverse primer amplification product; (c) denaturing the reverse primer amplification product from its template; (d) hybridizing a forward primer to: (e) extending the forward primer using the complementary target nucleic acid molecule, if present, or the reverse primer amplification product, as a template, wherein the forward primer joined to the forward primer extension product constitutes a forward primer amplification product; (f) denaturing the forward primer amplification product from its template; (g) hybridizing the reverse primer to the forward primer amplification product; (h) extending the reverse primer using the forward primer amplification product as a template to form an additional reverse primer extension product, wherein the reverse primer joined to the additional reverse primer extension product constitutes an additional reverse primer amplification product; (i) denaturing the additional reverse primer amplification product from its template; (j) hybridizing the forward primer to the reverse primer amplification product; (k) extending the forward primer using the reverse primer amplification product as a template to form an additional forward primer extension product, wherein the forward primer joined to the additional forward primer extension product constitutes an additional forward primer amplification product; (l) denaturing the additional forward primer amplification product from its template; (m) repeating steps (g) through (I), using the additional reverse primer amplification product and the additional forward primer amplification product as templates for the forward primer and the reverse primer, respectively, a sufficient number of times to produce a detectable quantity of additional reverse primer amplification product or of additional forward primer amplification product; and (n) detecting the presence of the additional reverse primer amplification product or the additional forward primer amplification product; the improvement wherein the nucleotide at the 3′ end of the reverse primer hybridizes with: wherein the nucleotide at the 3′ end of the forward primer hybridizes with: