rapid diagnosis of enterovirus infections by real-time pcr on the lightcycler using the taqman format论文.pdf
Diagnostic Microbiology and Infectious Disease 42 (2002) 99–105 Virology Rapid diagnosis of enterovirus infections by real-time PCR on the LightCycler using the TaqMan format Thomas Watkins-Riedela,*, Markus Woegerbauerb, David Hollemanna, Peter Hufnaglc aThe Division of Clinical Virology, Institute of Virology and Infectious Diseases, University Hospital Vienna, A-1090 Vienna, Austria bDepartment of Internal Medicine, University Hospital Vienna, A-1090 Vienna, Austria cRoche Diagnostics GmbH, A-1211 Vienna, Austria Received 17 July 2001; accepted 27 October 2001 Abstract 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
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