Detection of Foot-and-Mouth Disease Virus: Comparative Diagnostic Sensitivity of Two Independent Real-Time Reverse Transcription-Polymerase Chain Reaction Assays

King, Donald P.; Ferris, N.P.; Shaw, Andrew; Reid, Scott M.; Hutchings, G.; Giuffre, A.; Robida, John M.; Callahan, Johnny D.; Nelson, William M.; Beckham, Tammy R.

doi:10.1177/104063870601800114

Cited by 84 publications

(68 citation statements)

References 10 publications

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“…The development of an accurate and sensitive method for detection of FMDV required the recognition of conserved sequence in FMDV genome, located in all serotypes of FMDV and less affected by mutations such as 2B, 3D, 5UTR (IRES) regions (Vangrysperre and de Clercq, 1996;King et al, 2006). In this study, we used IRES and 3D regions as targets to perform an assay of sensitive, fast and accurate detection of FMDV.…”

Section: Discussionmentioning

confidence: 99%

Rapid Molecular Detection of Genetically Diverted Foot and Mouth Disease Virus Serotype O During the Outbreak of 2012 in Egypt

Rahman¹,

Hoffmann²,

Haas³

et al. 2015

International J. of Virology

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Increasing the international trade of animals and their products and the continuous changing of Foot and Mouth Disease Virus (FMDV) lead to the rise of disease introductions and create an urgent need for laboratory methods facilitating a swift and sensitive confirmation of suspect outbreaks and fast characterization of new isolates. As FMDV is extremely contagious and affects all cloven-hoofed animals, it provides a continuous burden and risk to the livestock industries of the developing world, in particular due to mortality in young animals, weight and milk loss, lameness and the trade restrictions necessary to control the disease. The control of FMD in Egypt requires an accurate analysis of the newly introduced viral strains and an analysis of their relationship with the current circulating strains and the routinely used vaccines. This study evaluates a recently developed rapid Reverse-Transcription Quantitative Polymerase Chain Reaction (RT-qPCR) for the identification of FMDV in samples from suspect cases. Samples were collected from gum, tongue epithelium, vesicular fluid, buccal and gum swabs, as well as saliva of infected cattle, buffalo and sheep and examined using two RT-qPCR of routine and high-speed formats using "IRES1" and "3D-OIE" primers. In addition, partial VP1 sequencing of some selected isolates was carried out for phylogenetic analysis. The high-speed RT-qPCR allowed a reliable diagnosis of FMDV in less than half an hour and can be used as a fast and valuable method for the monitoring and controlling of the foot and mouth disease. A new variant genotype (FMD-O/Eg/Mans/14) was circulating in Egypt during the outbreak of 2012 showing 98.5% identity to the isolated strain in Sudan (SUD_6_2008).

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Section: Discussionmentioning

confidence: 99%

Rapid Molecular Detection of Genetically Diverted Foot and Mouth Disease Virus Serotype O During the Outbreak of 2012 in Egypt

Rahman¹,

Hoffmann²,

Haas³

et al. 2015

International J. of Virology

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“…The performance of mRT-PCR and singleplex rRT-PCRs was compared using VI and Ag-ELISA to define FMDV true positives and true negatives. Although VI and Ag-ELISA are established methods for the detection of FMDV, previous studies (14,35) have demonstrated that rRT-PCR has higher diagnostic sensitivity and can detect virus in additional samples, which for the purposes of this study would be classified as "true" negatives. For the majority of true positive field samples, the 3D mRT-PCR response was saturated and grouped far from the cut-off.…”

Section: Discussionmentioning

confidence: 99%

“…Total nucleic acid was extracted from each ES by an automated procedure using a MagNA Pure LC (Roche, UK) as previously described (14,35). Extracted samples (40 µL) were aliquoted (3×13 µL), stored at -80ºC and thawed once just before use.…”

Section: Nucleic Acid Extractionmentioning

confidence: 99%

“…The 5´UTR and 3D rRT-PCR assays were initially compared prior to their implementation in Australia (3). A subsequent in-depth comparative evaluation (14) was conducted to further evaluate the effectiveness of these assays; demonstrating a higher diagnostic sensitivity of the rRT-PCR assays over VI and/or antigen-ELISA (35), particularly when both assays were used in combination. Both assays are used routinely in combination at the Food and Agriculture Organization of the United Nations, World…”

Section: Introductionmentioning

confidence: 99%

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Diagnostic evaluation of a multiplexed RT-PCR microsphere array assay for the detection of foot-and-mouth and look-alike disease viruses

Hindson¹,

Baker²,

Tammero³

et al. 2007

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A high-throughput multiplexed assay (Multiplex Version 1.0) was developed for the differential laboratory diagnosis of foot-and-mouth disease virus (FMDV) from viruses which cause clinically similar diseases of livestock. This assay simultaneously screens for five RNA and two DNA viruses using multiplexed reverse transcription PCR (mRT-PCR) amplification coupled with a microsphere hybridization array and flow-cytometric detection. Two of the seventeen primer-probe sets included in this multiplex assay were adopted from previously characterized

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“…An in-depth comparative evaluation of diagnostic sensitivity (D-SN) 42 was reported in which samples of epithelial suspension from tissue collected from suspect FMD cases were analyzed. In combination, these singleplex Taqman assays reported higher D-SN compare to virus isolation and antigen-ELISA.…”

Section: Foot-and-mouth Disease Virus (Fmdv) -Bovine/porcine Panelsmentioning

confidence: 99%

Development and Characterization of A Multiplexed RT-PCR Species Specific Assay for Bovine and one for Porcine Foot-and-Mouth Disease Virus Rule-Out

Smith¹,

Danganan²,

Tammero³

et al. 2007

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, 925-422-5545, smith350@llnl.gov Contributors:Linda Danganan, 925-422-2553, danganan1@llnl.gov Lance Tammero, PhD., 925-423-9545, tammero2@llnl.gov Beth Vitalis, PhD., 925-422-0149, vitalis1@llnl.gov Chemical and Biological Countermeasures Division Nonproliferation, Homeland and International Security (NHI)Lawrence Livermore National Laboratory (LLNL) Livermore, CA Principle Investigators: Ray Lenhoff, Ph.D., PhD., 925.422.5665, pnaraghi@llnl.gov Benjamin Hindson, PhD., hindson1@llnl.gov This work was performed under the auspices of the U.S. . This effort has the ability to improve our nation's capability to discriminate between foreign animal diseases and those that are endemic using a single assay, thereby increasing our ability to protect food and agricultural resources with a diagnostic test which could enhance the nation's capabilities for early detection of a foreign animal disease.In FY2005 A timeline for this development is presented in Table 1. The development of the Version 1.0 panel for FMDV rule-out and the most current efforts aimed to designed species specific panels has spanned over 2 ½ years with multiple collaborative partnerships.This document provides a summary of the development, testing and performance data at OIE Stage 1 Feasibility into Stage 2 Assay Development and Standardization 1 (see Table 2), gathered as of June 30 th , 2007 for the porcine and bovine MUX assay panels. We present an overview of the identification and selection of candidate genetic signatures, the assay development process, and preliminary performance data for each of the individual signatures as characterized in the multiplexed format for the porcine and bovine panels. The Stage 1 Feasibility data of the multiplexed panels is presented in this report also includes relevant data acquired from the Version 1.0 panel as supporting information where appropriate. In contrast to last years' effort, the development of the bovine and porcine panels is pending additional work to complete analytical characterization of FMDV, VESV, SVD, RPV and MCF. The signature screening process and final panel composition impacts this effort. The unique challenge presented this year was having strict predecessor limitations in completing characterization, where efforts at LLNL must precede efforts at PIADC, such challenges were alleviated in the 2006 reporting by having characterization data from the interlaboratory comparison and at Plum Island under AgDDAP project. We will present an addendum at a later date with additional data on the characterization of the porcine and bovine multiplex assays when that data is available. As a summary report, this document does not provide the details of 90% complete: during development, each MUX panel was tested against viral nucleic acid extracts (total nucleic acids, non-titered) from each organism. Preliminary results assessed by low-resolution titration curves, for each step-wise addition of assay to the multiplexed panel. 1.2.1.2.Calibration test of reference standards 90% complete: refere...

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Detection of Foot-and-Mouth Disease Virus: Comparative Diagnostic Sensitivity of Two Independent Real-Time Reverse Transcription-Polymerase Chain Reaction Assays

Cited by 84 publications

References 10 publications

Rapid Molecular Detection of Genetically Diverted Foot and Mouth Disease Virus Serotype O During the Outbreak of 2012 in Egypt

Rapid Molecular Detection of Genetically Diverted Foot and Mouth Disease Virus Serotype O During the Outbreak of 2012 in Egypt

Diagnostic evaluation of a multiplexed RT-PCR microsphere array assay for the detection of foot-and-mouth and look-alike disease viruses

Development and Characterization of A Multiplexed RT-PCR Species Specific Assay for Bovine and one for Porcine Foot-and-Mouth Disease Virus Rule-Out

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