Rapid diagnosis of enterovirus infections by real-time PCR on the LightCycler using the TaqMan format

A 2 h single-tube, reverse-transcription(RT)-PCR/hybridization assay using the TaqMan format for rapid diagnostic screening of enterovirus (EV) infections was optimized for the real-time LightCycler (LC) technology. For low EV load clinical samples an additional 30 min reamplification step using a n...

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Bibliographic Details
Published in:Diagnostic microbiology and infectious disease 2002-02, Vol.42 (2), p.99-105
Main Authors: Watkins-Riedel, Thomas, Woegerbauer, Markus, Hollemann, David, Hufnagl, Peter
Format: Article
Language:English
Subjects:
Antibiotics. Antiinfectious agents. Antiparasitic agents
Antifungal agents
Bacteriological methods and techniques used in bacteriology
Bacteriology
Biological and medical sciences
Enterovirus - genetics
Enterovirus - isolation & purification
Enterovirus Infections - diagnosis
Enterovirus Infections - virology
Feces - virology
Fluorescent Dyes
Fundamental and applied biological sciences. Psychology
Humans
Medical sciences
Microbiology
Pharmacology. Drug treatments
Pharynx - virology
Polymerase Chain Reaction - methods
Reverse Transcriptase Polymerase Chain Reaction
RNA, Viral - analysis
RNA, Viral - blood
RNA, Viral - cerebrospinal fluid
Sensitivity and Specificity
Taq Polymerase - metabolism
Time Factors
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Summary:A 2 h single-tube, reverse-transcription(RT)-PCR/hybridization assay using the TaqMan format for rapid diagnostic screening of enterovirus (EV) infections was optimized for the real-time LightCycler (LC) technology. For low EV load clinical samples an additional 30 min reamplification step using a novel primer-mix/probe design resulted in a 100% concordance with AMPLICOR EV PCR Test and in-house RT-PCR. Combined with maximum specificity, the sensitivity of LC-PCR was 10- to 100-fold higher compared to AMPLICOR EV Test.
ISSN:0732-8893
1879-0070
DOI:10.1016/S0732-8893(01)00330-3