| US 7,476,520 B2 | ||
| Primers for use in detecting beta-lactamases | ||
| Nancy D. Hanson, Gretna, Nebr. (US); Christine C. Sanders, Englewood, Fla. (US); and Anton F. Ehrhardt, Omaha, Nebr. (US) | ||
| Assigned to Creighton University, Omaha, Nebr. (US) | ||
| Filed on Mar. 21, 2005, as Appl. No. 11/84,973. | ||
| Application 09/814252 is a division of application No. 09/407818, filed on Sep. 28, 1999, granted, now 6,242,223, filed on Jun. 05, 2001. | ||
| Application 11/084973 is a continuation of application No. 09/814252, filed on Mar. 21, 2001, granted, now 6,893,846, filed on May 17, 2005. | ||
| Claims priority of provisional application 60/102181, filed on Sep. 28, 1998. | ||
| Claims priority of provisional application 60/121765, filed on Feb. 26, 1999. | ||
| Prior Publication US 2005/0260633 A1, Nov. 24, 2005 | ||
| Int. Cl. C12P 19/34 (2006.01); C12Q 1/68 (2006.01); C07H 21/02 (2006.01); C07H 21/04 (2006.01); C07H 21/00 (2006.01) | ||
| U.S. Cl. 435—91.2 [435/6; 435/91.1; 536/23.1; 536/24.3; 536/24.33; 536/25.3] | 5 Claims |
| 1. A method for analyzing a beta-lactamase of the SHV family of beta-lactamases in a clinical sample, the method comprising:
providing a clinical sample suspected of containing a beta-lactamase nucleic acid of the SHV family of beta-lactamases;
providing a pair of oligonucleotide primers, wherein the primers specifically hybridize with a beta-lactamase nucleic acid
of the SHV family of beta-lactamases, wherein one primer of the pair is complementary to at least a portion of the sense strand
of the beta-lactamase nucleic acid and the other primer of the pair is complementary to at least a portion of the antisense
strand of the beta-lactamase nucleic acid, and wherein at least one of the primers is selected from the group consisting of:
5′-ATC GTC CAC CAT CCA CTG CA-3′ (SEQ ID NO:6);
5′-TAG TGG ATC TTT CGC TCC AG-3′ (SEQ ID NO:8);
5′-CAC TCA AGG ATG TAT TGT G-3′ (SEQ ID NO:10); and full-length complements thereof;
annealing the primers to the beta-lactamase nucleic acid of the SHV family of the beta-lactamases if the beta-lactamase nucleic
acid of the SHV family of beta-lactamases is in the sample;
simultaneously extending the annealed primers from a 3′ terminus of each primer of the pair and synthesizing extension products
that are complementary to the nucleic acid strands of the beta-lactamase nucleic acid annealed to each primer of the pair
wherein the extension products after separation from one strand of the beta-lactamase nucleic acid serve as templates for
the synthesis of extension products of other strand of the beta-lactamase nucleic acid;
separating the extension products; and
analyzing the separated extension products for a region characteristic of the beta-lactamase of the SHV family if the beta-lactamase
nucleic acid of the SHV family of beta-lactamases is in the sample.
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