Improvement of a real-time RT-PCR assay for the detection of enterovirus RNA.
We describe an improvement of an earlier reported real-time RT-PCR assay for the detection of enterovirus RNA, based on the 5' exonuclease digestion of a dual-labeled fluorogenic probe by Taq DNA polymerase. A different extraction method, real-time RT-PCR instrument and primer set were evaluated. Our data show that the optimized assay yields a higher sensitivity and reproducibility and resulted in a significant reduced hands-on time per sample.
Piqueur
Marian A C
MA
Department of Microbiology, ZNA Hospitals, site Middelheim, Lindendreef 1, 2020 Antwerp, Belgium. marianpiqueur@hotmail.com
Verstrepen
Walter A
WA
Bruynseels
Peggy
P
Mertens
An H
AH
D017931:DNA Primers / Q000235:genetics; D004770:Enterovirus / Q000302:isolation & purification*; D004769:Enterovirus Infections / Q000175:diagnosis*; D006801:Humans; D012367:RNA, Viral / Q000302:isolation & purification*; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000295:instrumentation / Q000379:methods*; D012680:Sensitivity and Specificity
D017931:DNA Primers; D012367:RNA, Viral