Use of a portable real-time reverse transcriptasepolymerase chain reaction assay for rapid detection of foot-and-mouth disease virus

Callahan, Johnny D.; Brown, F.; Osorio, Fernando A.; Sur, Jung-Hyang; Kramer, Ed; Long, Gary W.; Liu, Juan; Ellis, Stefanie J; Shoulars, Katina S; Gaffney, Kristin L; Rock, Daniel L.; Nelson, William M.

doi:10.2460/javma.2002.220.1636

Cited by 360 publications

(372 citation statements)

References 12 publications

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“…All seven serotypes of FMDV grown in tissue culture were detected with viral RNA detectable 24-96 h before development of clinical signs. The efficiency of the assay in this application was evident when test results were available within two hours (Callahan et al, 2002).…”

Section: Comparison Of Methodsmentioning

confidence: 98%

“…The assay is also efficient since use of test plates specifically designed for the iCycler iQ ™ allowed assay of up to 384 samples/plate. The Weinstock, 2001 andMahlum et al, 2002), foot and mouth disease virus (Callahan et al, 2002), porcine endogenous retrovirus (Argaw et al, 2002), and rotavirus (Schwarz et al, 2002). Real-time RT-PCR assays were shown to be most efficient and sensitive for assay of clinical specimens infected with BVDV, when compared to gel-based RT-PCR assays, virus isolation tests, immunoperoxidase monolayer assays, and immunohistochemistry assays.…”

Section: Comparison Of Methodsmentioning

confidence: 99%

See 1 more Smart Citation

Detection and quantitation of bovine respiratory syncytial virus using real-time quantitative RT-PCR and quantitative competitive RT-PCR assays

Achenbach

Topliff

Vassilev

et al. 2004

Journal of Virological Methods

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Section: Comparison Of Methodsmentioning

confidence: 98%

Section: Comparison Of Methodsmentioning

confidence: 99%

Detection and quantitation of bovine respiratory syncytial virus using real-time quantitative RT-PCR and quantitative competitive RT-PCR assays

Achenbach

Topliff

Vassilev

et al. 2004

Journal of Virological Methods

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“…Detection of human immunodeficiency virus antibodies in oral fluids is currently being applied for clinical use and epidemiological surveillance 11 . In pigs, foot‐and‐mouth disease virus, Erysipelothrix species, influenza virus, porcine reproductive and respiratory syndrome virus (PRRSV), and porcine circovirus type 2 have been detected in oral fluids from naturally or experimentally infected pigs 1 , 2 , 12 , 13 …”

Section: Introductionmentioning

confidence: 99%

Sensitivity of oral fluids for detecting influenza A virus in populations of vaccinated and non‐vaccinated pigs

Romagosa

Gramer

Joo

et al. 2011

Influenza Resp Viruses

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Please cite this paper as: Romagosa et al. (2011) Sensitivity of oral fluids for detecting influenza A virus in populations of vaccinated and non‐vaccinated pigs. Influenza and Other Respiratory Viruses. Background/objective We evaluated the sensitivity of PCR on oral fluids in detecting influenza virus in vaccinated and non‐vaccinated pigs. Methods Three‐week‐old influenza‐free pigs were divided into three groups: (i) control, non‐vaccinated, (ii) vaccinated with a commercial, heterologous vaccine, and (iii) vaccinated with an experimental, homologous vaccine. After vaccination, an influenza‐infected pig was placed in contact with each of the groups. Individual nasal swabs and pen oral fluids were collected daily. Viral RNA was tested for the presence of influenza by RRT‐PCR and virus isolation attempted from oral fluids. A pen was considered positive if at least one nasal swab was positive. Results Based on nasal swab results, 43·8% of pens were detected positive but only 35% based on oral fluids. Overall sensitivity of oral fluids was 80%, and virus was isolated from 51% of RRT‐PCR‐positive oral fluids. The kappa coefficient for agreement (ĸ) between oral fluids and nasal swabs was 0·82. Among groups, ĸ was 1 (95% CI, 1–1), 0·74 (95% CI, 0·55–0·92), and 0·76 (95% CI, 0·5–1) for control, heterologous, and homologous‐vaccinated groups, respectively. There was less agreement when within pen prevalence was 10% or less. Probability of detecting influenza virus in oral fluids was 99% when within pen prevalence was higher than 18% and decreased to 69% when prevalence was 9%. Conclusions Results indicated that pen‐based collection of oral fluids is a sensitive method to detect influenza even when within pen prevalence is low and when pigs have been vaccinated and highlight the potential use of oral fluids for influenza surveillance.

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“…5'-CCYRGTCCCCTTCTCAGAT-3' 10.00 FMD-IRES1-3FAM 5'-FAM-CCCTTCAGGTACCCCGA GGTAACA-BHQ1-3' 2.5 Callahan et al (2002) FMD-3D-OIE FMD-3D-OIE-F 5'-ACTGGGTTTTACAAACCTGTGA-3' 10.00 FMD-3D-OIE-R 5'-GCGAGTCCTGCCACGGA-3' 10.00…”

Section: Routine and High-speed Rt-qpcr Analysismentioning

confidence: 99%

“…The FMD-IRES1 and FMD-3D-OIE primers were selected for detecting the IRES and the3D (polymerase RNA gene) fragments respectively (Callahan et al, 2002;Wernike et al, 2013). The used probes were labeled with 6-carboxyfluorescein (FAM) at the 5' end and the quencher tetramethylrhodamine (TAMRA) at the 3' end.…”

Section: Extraction Of Viral Rnamentioning

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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Use of a portable real-time reverse transcriptasepolymerase chain reaction assay for rapid detection of foot-and-mouth disease virus

Abstract: The assay reagents are produced in a vitrified form, which permits storage and transportation at ambient temperatures. The test can be performed in 2 hours or less on a portable instrument, thus providing a rapid, portable, sensitive, and specific method for detection of FMDV.

Cited by 360 publications

References 12 publications

Detection and quantitation of bovine respiratory syncytial virus using real-time quantitative RT-PCR and quantitative competitive RT-PCR assays

Detection and quantitation of bovine respiratory syncytial virus using real-time quantitative RT-PCR and quantitative competitive RT-PCR assays

Sensitivity of oral fluids for detecting influenza A virus in populations of vaccinated and non‐vaccinated pigs

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

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