Abstract
Highly sensitive and specific nucleic acid amplification tests (NAATs) have emerged as the gold standard diagnostic tests for many infectious diseases. Real-time PCR has further refined the technology of nucleic acid amplification with detection in a closed system and enabled multiplexing to simultaneously detect multiple pathogens. It is a versatile, fast, and high-throughput system for pathogen detection that has reduced the risk of PCR contamination, eliminated post-PCR manipulations, and improved the cost-effectiveness of testing. In addition, real-time PCR can be applied to self-collected noninvasive specimens. Here, we describe an in-house developed TaqMan-based real-time multiplex PCR (M-PCR) assay for the diagnosis of sexually transmitted genital ulcer disease (GUD) and discuss briefly on issues associated with validation of assay performance.
MeSH terms
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DNA, Bacterial / genetics
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DNA, Bacterial / isolation & purification
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DNA, Viral / genetics
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DNA, Viral / isolation & purification
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Haemophilus ducreyi / genetics
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Haemophilus ducreyi / isolation & purification
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Haemophilus ducreyi / pathogenicity
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Herpesvirus 1, Human / genetics
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Herpesvirus 1, Human / isolation & purification
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Herpesvirus 1, Human / pathogenicity
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Herpesvirus 2, Human / genetics
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Herpesvirus 2, Human / isolation & purification
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Herpesvirus 2, Human / pathogenicity
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Humans
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Molecular Diagnostic Techniques / methods*
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Real-Time Polymerase Chain Reaction
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Ribonuclease P / genetics
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Sexually Transmitted Diseases / diagnosis*
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Sexually Transmitted Diseases / genetics
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Sexually Transmitted Diseases / microbiology
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Sexually Transmitted Diseases / virology
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Taq Polymerase / metabolism
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Treponema pallidum / genetics
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Treponema pallidum / isolation & purification
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Treponema pallidum / pathogenicity
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Ulcer / diagnosis*
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Ulcer / genetics
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Ulcer / microbiology
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Ulcer / virology
Substances
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DNA, Bacterial
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DNA, Viral
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Taq Polymerase
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Ribonuclease P