Polynucleotides for the amplification and detection of chlamydia trachomatis and neisseria gonorrhoeae

Abstract

Polynucleotides useful for detecting Chlamydia trachomatis and/or Neisseria gonorrhoeae in a test sample, kits, a nucleic acid amplification method and detection method including the same.

What is claimed is: 
     
         1 . A composition comprising two or more polynucleotides having nucleic acid sequences selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, and SEQ ID NO:12. 
     
     
         2 . The composition of  claim 1  comprising a first polynucleotide having the nucleic acid sequence of SEQ ID NO: 1 or SEQ ID NO: 2 and a second polynucleotide having the nucleic acid sequence of SEQ ID NO: 4. 
     
     
         3 . The composition of  claim 1  comprising a first polynucleotide having the nucleic acid sequence of SEQ ID NO: 8 and a second polynucleotide having the nucleic acid sequence of SEQ ID NO: 9. 
     
     
         4 . The composition of  claim 1  comprising a first polynucleotide having the nucleic acid sequence of SEQ ID NO: 2, a second polynucleotide having the nucleic acid sequence of SEQ ID NO: 4, a third polynucleotide having the nucleic acid sequence of SEQ ID NO: 8 and a fourth polynucleotide having the nucleic acid sequence of SEQ ID NO: 9. 
     
     
         5 . A primer/probe set selected from the primer/probe sets consisting of:
 Primer and Probe Set 1 (SEQ ID NOs: 1, 4, and 5);   Primer and Probe Set 2 (SEQ ID NOs: 2, 4, and 5);   Primer and Probe Set 3 (SEQ ID NOs: 3, 4, and 5);   Primer and Probe Set 4 (SEQ ID NOs: 1, 4, and 6);   Primer and Probe Set 5 (SEQ ID NOs: 2, 4, and 6);   Primer and Probe Set 6 (SEQ ID NOs: 3, 4, and 6);   Primer and Probe Set 7 (SEQ ID NOs: 1, 4, and 7);   Primer and Probe Set 8 (SEQ ID NOs: 2, 4, and 7);   Primer and Probe Set 9 (SEQ ID NOs: 3, 4, and 7);   Primer and Probe Set 10 (SEQ ID NOs: 8, 9, and 10);   Primer and Probe Set 11 (SEQ ID NOs: 8, 9, and 11); and   Primer and Probe Set 12 (SEQ ID NOs: 8, 9, and 12).   
     
     
         6 . The primer/probe set of  claim 5 , wherein the primer/probe set is
 Primer and Probe Set 4 (SEQ ID NOs: 1, 4, and 6).   
     
     
         7 . The primer/probe set of  claim 5 , wherein the primer/probe set is
 Primer and Probe Set 10 (SEQ ID NOs: 8, 9, and 10).   
     
     
         8 . A composition of matter comprising the primer/probe set of  claim 5 , wherein the primer/probe set is Primer and Probe Set 4 (SEQ ID NOs: 1, 4, and 6), further comprising primer/probe set 10 (SEQ ID NOs: 8, 9, and 10). 
     
     
         9 . A method of amplifying  Chlamydia trachomatis  and/or  Neisseria gonorrhoeae  in a test sample, said method comprising:
 (a) forming a reaction mixture comprising nucleic acid amplification reagents, a test sample potentially containing a  Chlamydia trachomatis  and/or  Neisseria gonorrhoeae  target sequence a composition according to  claim 1 ; and   (b) subjecting the mixture to amplification conditions to generate at least one copy of a nucleic acid sequence complementary to the target sequence.   
     
     
         10 . The method of  claim 9  wherein the amplification conditions include changing the temperature of the sample and the temperature change is repeated between 10 and 100 times. 
     
     
         11 . An isolated polynucleotide having a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 2, 4, 5, 6, 7, 8, 9, 10, 11, and 12. 
     
     
         12 . The isolated polynucleotide of  claim 11 , comprising a detectable label. 
     
     
         13 . A method of detecting  Chlamydia trachomatis  and/or  Neisseria gonorrhoeae  in a test sample, said method comprising:
 (a) forming a reaction mixture comprising nucleic acid amplification reagents, a test sample potentially containing a  Chlamydia trachomatis  and/or  Neisseria gonorrhoeae  target sequence, and
 (i) at least one polynucleotide according to  claim 11 , or 
 (ii) at least one composition according to  claim 1  or 
 (iii) at least one primer/probe set according to  claim 5 ; 
   (b) subjecting the mixture to amplification conditions to generate at least one copy of a nucleic acid sequence complementary to the target sequence;   (c) hybridizing the probe to the nucleic acid sequence complementary to the target sequence, so as to form a hybrid comprising the probe and the nucleic acid sequence complementary to the target sequence; and   (d) detecting the hybrid as an indication of the presence of  Chlamydia trachomatis  and/or  Neisseria gonorrhoeae  in the test sample.   
     
     
         14 . The method of  claim 13  wherein said reaction mixture further comprises a control target polynucleotide and a control polynucleotide probe 
     
     
         15 . A kit comprising:
 (a) a polynucleotide of  claim 11 ; and   (b) amplification reagents.   
     
     
         16 . The kit of  claim 15  comprising:
 (a) two or more polynucleotides each having a distinct nucleic acid sequence selected from the group consisting of SEQ ID NOs: 2, 4, 5, 6, 7, 8, 9, 10, 11, and 12, and 
 (b) amplification reagents.