Evaluation of a new multiplex polymerase chain reaction assay STDFinder for the simultaneous detection of 7 sexually transmitted disease pathogens

CM Muvunyi, N Dhont, R Verhelst, T Crucitti, M Reijans, B Mulders, G Simons, M Temmerman, G Claeys, E Padalko

    Research output: Contribution to journalA1: Web of Science-article

    Abstract

    We evaluated a new multiplex polymerase chain reaction (mPCR), 'STDFinder assay', a novel multiplex ligation-dependent probe amplification (MLPA) assay for the simultaneous detection of 7 clinically relevant pathogens of STDs, i.e., Neisseria gonorrhoeae, Chlamydia trachomatis, Trichomonas vaginalis, Mycoplasma genitalium, Treponema pallidum, and herpes simplex virus type 1 and 2 (HSV-1 and HSV-2). An internal amplification control was included in the mPCR reaction. The limits of detection for the STDFinder assay varied among the 7 target organisms from 1 to 20 copies per MLPA assay. There were no cross-reactions among any of the probes. Two hundred and forty-two vaginal swabs and an additional 80 specimens with known results for N. gonorrhoeae and C. trachomatis, obtained from infertile women seen at an infertility research clinic at the Kigali Teaching Hospital in Rwanda, were tested by STDFinder assay and the results were confirmed by single real-time PCR using different species-specific targets. Compared to the reference standard, the STDFinder assay showed specificities and sensitivities of 100% and 100%, respectively, for N. gonorrhoeae, C. trachomatis, and M. genitalium; 90.2% and 100%, respectively, for Trichomonas vaginalis; and 96.1% and 100%, respectively, for HSV-2. No specimen was found to be positive for HSV-1 by either the STDFinder assay or the comparator method. Similarly, the sensitivity for Treponema pallidum could not be calculated due to the absence of any Treponema pallidum-positive samples. In conclusion, the STDFinder assays have comparable clinical sensitivity to the conventional mono and duplex real-time PCR assay and are suitable for the routine detection of a broad spectrum of these STDs at relatively low cost due to multiplexing.
    Original languageEnglish
    JournalDiagnostic Microbiology and Infectious Disease
    Volume71
    Issue number1
    Pages (from-to)29-37
    ISSN0732-8893
    DOIs
    Publication statusPublished - 2011

    Keywords

    • B780-tropical-medicine
    • Sexually transmitted diseases
    • Bacterial diseases
    • Gonorrhea
    • Neisseria gonorrhoeae
    • Chlamydia
    • Chlamydia trachomatis
    • Trichomoniasis
    • Trichomonas vaginalis
    • Mycoplasma genitalium
    • Syphilis
    • Treponema pallidum
    • Viral diseases
    • Herpes simplex virus type 2
    • HSV-2
    • HSV-1
    • Detection
    • Multiplex PCR
    • Assays
    • Amplification
    • Performance
    • Vaginal specimens
    • Women
    • Specificity
    • Sensitivity
    • Laboratory techniques and procedures

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