rapid diagnosis of enterovirus infections by real-time pcr on the lightcycler using the taqman format论文.pdf

文档介绍：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