Compositions and methods for detection of trichomonas vaginalis

• US 10,934,597 B2
• Filed: 05/22/2017
• Issued: 03/02/2021
• Est. Priority Date: 05/27/2016
• Status: Active Grant

• Current Assignee: Roche Molecular Systems (Roche Holding AG)
• Original Assignee: Roche Molecular Systems (Roche Holding AG)
• Inventors: Harris, Jody, Lu, Shi-Da Y., Mangipudi, Kalyani, Sun, Jingtao
• Primary Examiner(s): Goldberg, Jeanine A
• Application Number:
  US15/601,495
• Publication Number:
  US 20170342508A1
• Time in Patent Office: 1,380 Days
• Field of Search: None
• CPC Class Codes: C12Q 1/689 for bacteria
• CPC Class Codes: C12Q 1/6893 for protozoa
• CPC Class Codes: C12Q 2600/158 Expression markers

Abstract

Methods for the rapid detection of the presence or absence of Trichomonas vaginalis (TV) in a biological or non-biological sample are described. The methods can include performing an amplifying step, a hybridizing step, and a detecting step. Furthermore, primers, probes targeting the target TV gene, along with kits are provided that are designed for the detection of TV.

Claims

1. A method of specifically amplifying and detecting Trichomonas vaginalis (TV) in a sample by discriminating against Trichomonas tenax and Pentatrichomonas homini the method comprising:

performing an amplifying step comprising contacting the sample with a set of target TV gene primers to produce an amplification product if a target TV gene nucleic acid is present in the sample, wherein the target TV gene is the 5.8s ribosomal RNA gene;

performing a hybridizing step comprising contacting the amplification product with one or more detectable target TV gene probes; and

detecting the presence or absence of the amplification product, wherein the presence of the amplification product is indicative of the presence of TV in the sample and wherein the absence of the amplification product is indicative of the absence of TV in the sample;

wherein the set of target TV gene primers comprise a first primer consisting of SEQ ID NO;

3 and a second primer consisting of SEQ ID NO;

12; and

wherein the detectable target TV gene probes comprises a third oligonucleotide sequence consisting of SEQ ID NO;

18;

and wherein the method does not detect Trichomonas tenax and Pentatrichomonas hominis.

2. The method of claim 1, wherein:

the hybridizing step comprises contacting the amplification product with the detectable target TV gene probe that is labeled with a donor fluorescent moiety and a corresponding acceptor moiety; and

the detecting step comprises detecting the presence or absence of fluorescence resonance energy transfer (FRET) between the donor fluorescent moiety and the acceptor moiety of the probe, wherein the presence or absence of fluorescence is indicative of the presence or absence of TV in the sample.

3. The method of claim 2, wherein said amplification step employs a polymerase enzyme having 5′

to 3′

nuclease activity.

4. The method of claim 2, wherein the donor fluorescent moiety and the corresponding acceptor moiety are within no more than 8-20 nucleotides of each other on the probe.

5. The method of claim 2, wherein the acceptor moiety is a quencher.