Polynucleotides for the amplification and detection of
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.
1. A method of amplifying Chlamydia trachomatis 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 target sequence and a composition of 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, and SEQ ID NO: 7, wherein one of the at least two or more polynucleotides comprises SEQ ID NO: 3; and
(b) subjecting the mixture to amplification conditions to generate at least one copy of a nucleic acid sequence complementary to the target sequence.
2. The method of claim 1 wherein the amplification conditions include changing the temperature of the sample and the temperature change is repeated between 10 and 100 times.
3. A method of detecting Chlamydia trachomatis 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 target sequence, and
(i) at least one composition of 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, and SEQ ID NO: 7, wherein one of the at least two or more polynucleotides comprises SEQ ID NO: 3, or
(iii) at least one primer/probe set consisting of:
Primer and Probe Set 3 (SEQ ID NOs: 3, 4, and 5);
Primer and Probe Set 6 (SEQ ID NOs: 3, 4, and 6);
Primer and Probe Set 9 (SEQ ID NOs: 3, 4, and 7);
(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 in the test sample.
4. The method of claim 3 wherein said reaction mixture further comprises a control target polynucleotide and a control polynucleotide probe.