Metronidazole resistance in trichomonas vaginalis and single nucleotide polymorphisms
The present invention is directed to the discovery of single nucleotide polymorphisms (SNPs) in the presence of metronidazole-resistant Trichomonas vaginalis . The presence of G76C, C213G, or C318A (SNP) in tvntr 4 or the presence of A238T, G427C, or T476C (SNP) in tvntr6 provides a reliable biomarker for the presence of metronidazole-resistant Trichomonas vaginalis . The present invention further provides reagents used for detecting the SNPs to screen subjects for metronidazole resistance in Trichomonas vaginalis.
1. A kit for detecting metronidazole resistance in Trichomonas vaginalis in a human, comprising:
(a) one or a plurality of reagents for detecting the presence of A238T in tvntr6 in Trichomonas vaginalis present in a human, wherein the reagent or plurality of reagents comprises a fluorescently labeled probe consisting of SEQ ID NO: 7; and
(b) an instruction for use of the reagent or a plurality of reagents in detecting the presence of A238T in trntr6 in Trichomonas vaginalis , wherein the presence of A238T in trntr6 in Trichomonas vaginalis is indicative metronidazole resistance in Trichomonas vaginalis.
2. The kit of claim 1 , wherein the plurality of reagents comprises a forward primer consisting of SEQ ID NO 5 and a reverse primer consisting of SEQ ID NO 6.
3. The kit of claim 1 , wherein the plurality of reagents comprises a probe consisting of SEQ ID NO: 11, a forward primer consisting of SEQ ID NO: 9 and a reverse primer consisting of SEQ ID NO: 10.
4. The kit of claim 1 further comprising a reagent for isolating nucleic acid.
5. The kit of claim 1 further comprising a reagent for allele-specific PCR and sequencing.
6. The kit of claim 1 further comprising a reagent for PCR and pyrosequencing.
7. The kit of claim 1 , wherein the plurality of reagents comprises a fluorescently labeled probe comprising a nucleic acid sequence consisting of SEQ ID NO 11.