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Compositions and methods for detection of trichomonas vaginalis

A technique for detecting Trichomonas vaginalis and samples, which is applied in the field of molecular diagnostics and can solve problems such as decreased sensitivity

Active Publication Date: 2019-01-04
F HOFFMANN LA ROCHE & CO AG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to convenience and relatively low cost, microscopic examination of wet preparations of genital secretions is probably the most commonly used method for the diagnosis of T. vaginalis, but is only 35% to 80% sensitive compared to culture
Furthermore, the sensitivity of the wet mount method is highly dependent on the experience of the microscopist and the time the specimen is transported to the laboratory, where the sensitivity drops by up to 20% within 1 hour of collection

Method used

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  • Compositions and methods for detection of trichomonas vaginalis
  • Compositions and methods for detection of trichomonas vaginalis
  • Compositions and methods for detection of trichomonas vaginalis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0153] Target selection for TV is a comprehensive search of public sequence databases as well as Trichomonas tenax and a literature search for potential TV targets of Trichomonas hominis. Multiple targets from public sequence databases were analyzed during target selection at the design stage, but all showed the same T. tenax Cross-reactivity with Pentatrimonas hominis. Sequences in public databases are complicated by "bulks" of sequence data from multi-copy targets. BLAST analysis of selected oligonucleotides indicated that the only significant cross-reactivity would be with Trichomonas tenax .

[0154] use cobas ® 4800 system or cobas ® The 6800 / 8800 system platform (Roche MolecularSystems, Inc., Pleasanton, CA) was used for real-time PCR detection of TV. The final concentrations of the amplification reagents are as follows:

[0155] Table XVI PCR amplification reagents

[0156]

[0157] The table below shows typical thermoprofiles for PCR amplification react...

Embodiment 2

[0165] Amplification and detection of the TV 5.8s rRNA gene was carried out as described in Example 1 (using primers with SEQ ID NO: 3 and 12 and a labeled probe with SEQ ID NO: 18), except that Mycoplasma genitalium Genomic template DNA of (MG) is included in the PCR assay along with primers and probes capable of amplifying and detecting MG. Primers and probes with SEQ ID NOs: 139, 145, and 150 that hybridize to the conserved region A of the mgpB gene (mgpB) and that hybridize to the variable region EF of the mgpB gene (MgPar) with SEQ ID NOs: Primers and probes for ID NOs: 153, 169 and 194.

[0166] The TV limit of detection (LOD) of 100, 10, 5, and 1 genome equivalent (ge / PCR) per PCR reaction was tested at a co-incubation with an internal control standard (GIC) and MG at 10 ge / PCR. carried out in amplification. The results are shown in image 3 (TV) and 4 (MG and GIC). All levels of TV were detected without dropout and the TV LOD was determined to be <1 ge / PCR. In add...

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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 for design of the detection of TV.

Description

field of invention [0001] The present disclosure relates to the field of molecular diagnostics, and more particularly to Trichomonas vaginalis (Trichomonas vaginalis) detection. Background of the invention [0002] Trichomonas vaginalis (TV) is a flagellated protozoan parasite that causes trichomoniasis, the most prevalent nonviral sexually transmitted infection in the United States, affecting approximately 3.7 million people nationwide. Prevalence differed between black and non-Hispanic white women, with 13% and 1.8% affected, respectively. Trichomonas vaginalis infection has been reported to affect >11% of women aged ≥40 years, and prevalence has been reported to be as high as 26% among symptomatic women tested in STD clinics and 6.5% among asymptomatic women. Trichomonas vaginalis is also known to cause urethritis in men who have sex with women (MSW). Most infections go unnoticed, with 70% of men and 85% of women experiencing only mild symptoms, which can persist ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/686C12Q1/6893
CPCC12Q1/686C12Q1/6893C12Q2537/143C12Q2565/1015
Inventor J.哈里斯S.D.Y.卢K.曼吉普迪J.孙
Owner F HOFFMANN LA ROCHE & CO AG
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