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 method of detecting metronidazole resistance in Trichomonas vaginalis in a human, comprising the steps of:
(a) obtaining a biological sample from a human;
(b) isolating DNA from said biological sample; and
(c) detecting the presence of a SNP selected from the group consisting of G76C, C213G and C318A in tvntr4, or a SNP selected from the group consisting of A238T, G427C and T476C in tvntr6 in said DNA,
wherein the presence of said SNP in said biological sample is indicative of metronidazole resistance in Trichomonas vaginalis present in said biological sample.
2. The method of claim 1 , wherein said SNP is G76C in tvntr4.
3. The method of claim 1 , wherein said SNP is C213G in tvntr4.
4. The method of claim 1 , wherein said SNP is C318A in tvntr4.
5. The method of claim 1 , wherein said SNP is A238T in tvntr6.
6. The method of claim 1 , wherein said SNP is G427C in tvntr6.
7. The method of claim 1 , wherein said SNP is T476C in tvntr6.
8. The method of claim 1 , wherein said biological sample is a vaginal swab or cervico-vaginal swab.
9. The method of claim 1 , wherein said isolating step is performed using phenol-chloroform.
10. The method of claim 1 , wherein said detecting step is performed by polymerase chain reaction (PCR).
11. The method of claim 10 , wherein said PCR is a real-time PCR, a PCR followed by sequencing, a PCR followed by pyrosequencing, or an allele-specific PCR.