Detection of Vancomycin-Resistant Enterococcus spp.

Abstract

The invention provides methods to detect vancomycin-resistant enterococci in biological samples using real-time PCR. Primers and probes for the detection of vancomycin-resistant enterococci are provided by the invention. Articles of manufacture containing such primers and probes for detecting vancomycin-resistant enterococci are further provided by the invention.

1 . An article of manufacture, comprising: 
 a pair of vanA primers;    a pair of vanA probes; and    a donor fluorescent moiety and a corresponding fluorescent moiety.    
     
     
         2 . The article of manufacture of  claim 1 , wherein said pair of vanA primers comprises a first vanA primer and a second vanA primer, wherein said first vanA primer comprises the sequence 5′-CGA GGA CGG ATA CAG GA-3′ (SEQ ID NO:1), and wherein said second vanA primer comprises the sequence 5′-CTT ATC ACC CCT TTA ACG C-3′ (SEQ ID NO:2).  
     
     
         3 . The article of manufacture of  claim 1 , wherein said pair of vanA probes comprises a first vanA probe and a second vanA probe, wherein said first vanA probe comprises the sequence 5′-CAA GAT AAC GGC CGC ATT GTA CTG AAC GA-3′ (SEQ ID NO:3), and wherein said second vanA probe comprises the sequence 5′-GTC AAT ACT CTG CCC GGT TTC AC-3′ (SEQ ID NO:4).  
     
     
         4 . The article of manufacture of  claim 1 , wherein said pair of vanA probes comprises a first vanA probe labeled with said donor fluorescent moiety and a second vanA probe labeled with said corresponding acceptor fluorescent moiety.  
     
     
         5 . The article of manufacture of  claim 1 , further comprising a package label or package insert having instructions thereon for using said pair of vanA primers and said pair of vanA probes to detect the presence or absence of one or more vancomycin-resistant enterococci in a biological sample.  
     
     
         6 . An article of manufacture, comprising: 
 a pair of vanlb primers;    a pair of vanB probes; and    a donor fluorescent moiety and a corresponding fluorescent moiety.    
     
     
         7 . The article of manufacture of  claim 6 , wherein said pair of vanB primers comprises a first vanB primer and a second vanB primer, wherein said first vanB primer comprises the sequence 5′-GAA GAT ACC GTG GCT CA-3′ (SEQ ID NO:5), and wherein said second vanB primer comprises the sequence 5′-GTA CGG AAG AAC TTA ACG CT-3′ (SEQ ID NO:6).  
     
     
         8 . The article of manufacture of  claim 6 , wherein said pair of vanB probes comprises a first vanB probe and a second vanB probe, wherein said first vanB probe comprises the sequence 5′-GAT CCA CTT CGC CGA CAA-3′ (SEQ ID NO:7), and wherein said second vanB probe comprises the sequence 5′-AAA TCA TCC TCG TTT CCC AT-3′ (SEQ ID NO:8).  
     
     
         9 . The article of manufacture of  claim 6 , wherein said pair of vanB probes comprises a first vanB probe labeled with said donor fluorescent moiety and a second vanB probe labeled with said corresponding acceptor fluorescent moiety.  
     
     
         10 . The article of manufacture of  claim 6 , further comprising a package label or package insert having instructions thereon for using said pair of vanB primers and said pair of vanB probes to detect the presence or absence of vancomycin-resistant enterococci in a biological sample.