NDLI: Cross-platform evaluation of commercial real-time reverse transcription PCR master mix kits using a quantitative 5'nuclease assay for Ebola virus.

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Cross-platform evaluation of commercial real-time reverse transcription PCR master mix kits using a quantitative 5'nuclease assay for Ebola virus.

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Cross-platform evaluation of commercial real-time reverse transcription PCR master mix kits using a quantitative 5'nuclease assay for Ebola virus.

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	Stephens, Kenyatta W. Hutchins, Rebecca J. Dauphin, Leslie A.
Description	Country affiliation: United States Author Affiliation: Stephens KW ( Bioterrorism Rapid Response and Advanced Technology (BRRAT) Laboratory, Laboratory Preparedness and Response Branch (LPRB), Division of Preparedness and Emerging Infections (DPEI), Centers for Disease Control and Prevention (CDC), Mail Stop G-42, 1600 Clifton Road, Atlanta, GA 30333, USA.)
Abstract	Selection of optimal reaction master mix reagents is essential to obtain the best performance with diagnostic real-time reverse transcription polymerase chain reaction (RT-PCR) assays. Every year the number of commercially available master mix kits increases, so it is prudent to periodically evaluate kits on the market. In this study we evaluated five commercial real-time RT-PCR master mix kits, the RealMasterMix RT-PCR ROX kit, the AgPath-ID One-Step RT-PCR kit, the SuperScript III Platinum One-step Quantitative RT-PCR system, the QuantiTect Probe RT-PCR kit, and the LightCycler RNA HybProbe amplification kit, using a 5'nuclease assay for Ebola virus. The kits were evaluated using the manufacturer's recommended conditions, as well as conditions which have been used with the Ebola virus assay during outbreaks. When evaluated for use in Ebola virus RNA detection, the AgPath-ID kit resulted in the greatest sensitivity in comparison to the other four kits. The efficacy of the AgPath-ID kit was instrument-independent in the five real-time PCR platforms tested. This study demonstrated that Ebola virus RNA detection was not equivalent among the master mix reagents studied and, thus, that this variable can affect real-time RT-PCR assay sensitivity. Furthermore, this study rates the master mix reagents for their suitability, providing diagnostic laboratories the option to select from these kits to suit their specific laboratory needs for real-time RT-PCR.
File Format	HTM / HTML
ISSN	08908508
Issue Number	6
Volume Number	24
e-ISSN	10961194
Journal	Molecular and Cellular Probes
Language	English
Publisher	Elsevier
Publisher Date	2010-12-01
Publisher Place	Great Britain (UK)
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	Discipline Cell Biology Discipline Molecular Biology Discipline Biotechnology Deoxyribonucleases Metabolism Ebolavirus Genetics Enzyme Assays Methods Reagent Kits, Diagnostic Standards Reverse Transcriptase Polymerase Chain Reaction Animals Cercopithecus Aethiops Isolation & Purification Rna, Viral Analysis Economics Sensitivity And Specificity Time Factors Vero Cells Evaluation Studies Journal Article
Content Type	Text
Resource Type	Article
Subject	Cell Biology Molecular Biology

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4	Project PI / Joint PI	Principal Investigator and Joint Principal Investigators of the project	Dr. B. Sutradhar bsutra@ndl.gov.in Prof. Saswat Chakrabarti will be added soon
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