Assay for
A region of the Trichomonas vaginalis AP65-1 gene has been identified which is useful for performing amplification assays to determine specifically whether T. vaginalis is present in the sample being tested. Oligonucleotides useful for performing thermal Strand Displacement Assay (tSDA) reactions on this gene are disclosed. The disclosed oligonucleotides can be used in an assay which is specific for multiple strains of T. vaginalis and which does not show cross reactivity with the genomes of other microorganisms or with human DNA.
1. A PCR amplification assay for detecting the presence or absence of Trichomonas vaginalis , the assay comprising a set of a plurality of primers and a probe for the detection of the AP65-1 gene of Trichomonas vaginalis comprising a first oligonucleotide primer comprising SEQ ID NO: 12, a second oligonucleotide primer comprising SEQ ID NO: 13 and a probe comprising SEQ ID NO: 14 wherein the first and second oligonucleotide primers and the probe all hybridize within the SEQ ID NO: 24 portion of the AP65-1 gene and wherein the probe further comprises a detectable label attached to the probe oligonucleotide.
2. A kit for the amplification detection of Trichomonas vaginalis comprising:
a plurality of primers and at least one probe for the amplification detection of Trichomonas vaginalis that contains an AP65-1 gene (SEQ ID NO:1)
a) wherein the plurality of primers comprises a first oligonucleotide primer comprising SEQ ID NO: 12 and a second oligonucleotide primer comprising SEQ ID NO: 13 that bind to the highly conserved region identified as SEQ ID NO: 24; and
b) at least one probe comprising SEQ ID NO: 14 wherein the probe further comprises a detectable label attached to the probe oligonucleotide.
3. The kit of claim 2 wherein the nucleic acid amplification reaction is polymerase chain reaction (PCR), the kit further comprising a polymerase.