Cross-platform evaluation of commercial real-time SYBR green RT-PCR kits for sensitive and rapid detection of European bat Lyssavirus type 1

BioMed Research International
Evelyne Picard-MeyerFlorence Cliquet

Abstract

This study evaluates the performance of five two-step SYBR Green RT-qPCR kits and five one-step SYBR Green qRT-PCR kits using real-time PCR assays. Two real-time thermocyclers showing different throughput capacities were used. The analysed performance evaluation criteria included the generation of standard curve, reaction efficiency, analytical sensitivity, intra- and interassay repeatability as well as the costs and the practicability of kits, and thermocycling times. We found that the optimised one-step PCR assays had a higher detection sensitivity than the optimised two-step assays regardless of the machine used, while no difference was detected in reaction efficiency, R (2) values, and intra- and interreproducibility between the two methods. The limit of detection at the 95% confidence level varied between 15 to 981 copies/µL and 41 to 171 for one-step kits and two-step kits, respectively. Of the ten kits tested, the most efficient kit was the Quantitect SYBR Green qRT-PCR with a limit of detection at 95% of confidence of 20 and 22 copies/µL on the thermocyclers Rotor gene Q MDx and MX3005P, respectively. The study demonstrated the pivotal influence of the thermocycler on PCR performance for the detection of rabies RNA, as ...Continue Reading

References

May 27, 2003·Clinical Chemistry·Dagmar SchoderMartin Wagner
Aug 28, 2004·Clinical Chemistry·Anders StåhlbergMichael Pfaffl
Oct 7, 2004·Journal of Wildlife Diseases·Vidya ShankarThomas J O'shea
Jun 16, 2005·Journal of Clinical Microbiology·D SchoderM Wagner
Mar 1, 2006·Journal of Clinical Virology : the Official Publication of the Pan American Society for Clinical Virology·T NagarajV Ravi
May 15, 2008·BioTechniques·Young Ho KimSang-Ryoul Park
Jun 25, 2009·Journal of Medical Virology·Susan A Nadin-DavisAlexander I Wandeler
Sep 30, 2009·PLoS Neglected Tropical Diseases·Anthony R FooksThomas Müller
Sep 3, 2010·Journal of Clinical Microbiology·Jessica CoertseWanda Markotter
May 24, 2011·Journal of Virological Methods·Supaporn WacharapluesadeeThiravat Hemachudha
Jul 23, 2011·Journal of Virological Methods·David T S HaymanAnthony R Fooks
Apr 1, 2014·Journal of Virological Methods·Melina FischerBernd Hoffmann
Aug 6, 2014·Viruses·Ashley C BanyardAnthony R Fooks

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Citations

Feb 6, 2016·PLoS Neglected Tropical Diseases·Emmanuelle RobardetFlorence Cliquet
Oct 3, 2019·Tropical Medicine and Infectious Disease·Emmanuelle RobardetFlorence Cliquet
Aug 11, 2017·European Journal of Clinical Investigation·Stephen Bustin, Tania Nolan
Dec 14, 2018·Journal of Veterinary Diagnostic Investigation : Official Publication of the American Association of Veterinary Laboratory Diagnosticians, Inc·Evelyne Picard-MeyerFlorence Cliquet
Jul 9, 2020·Pharmacological Reports : PR·Máté Tamás DériKatalin Monostory
Nov 22, 2018·Zoonoses and Public Health·Evelyne Picard-MeyerFlorence Cliquet

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Methods Mentioned

BETA
PCR
PCRs
Assay

Software Mentioned

qScript
Perfecta
Rotor Gene Q Series
Mx3005Pro
QuantiTect
Maxima
Sso

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