This SOP describes a two-step protocol for PriProET real-time PCR amplification of swine vesicular disease viruses (SVDV). A key point of the assay is tolerance toward mutations in the probe region. This SOP describes a two-step protocol for PriProET real-time PCR amplification of swine vesicular disease viruses (SVDV). The assay amplifies the 3D-gene of any of SVDV strain while heterologous virus...

Authors: Mikhayil Hakhverdyan
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A simple and reliable technique using commercial reagents (TRIzol) to extract RNA from field samples. This report describes a simple and reliable extraction technique using commercial reagents (TRIzol) to provide RNA for subsequent reverse transcription and specific PCR amplification. Samples suitable for testing include field samples and cells suspected to contain RNA viruses of vesicular disease...

Authors: Own Document
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Sensitive and specific detection of Aujeszky’s disease virus (ADV), African swine fever virus (ASFV), porcine circovirus type 2 (PCV2) and porcine parvovirus (PPV). This protocol describes the sensitive and specific detection of Aujeszky’s disease virus (ADV), African swine fever virus (ASFV), porcine circovirus type 2 (PCV2) and porcine parvovirus (PPV). The assays target the gD gene (ADV), t...

Authors: Own Document
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The Invader™ assay is a linear, isothermal signal amplification system able to accurately quantify DNA and RNA targets with high sensitivity and specificity. This report describes the sensitive and specific detection of African swine fever virus DNA using the isothermal ASFV vp72 655T Invader squared assay. The Invader™ assay is a linear, isothermal (63?C) signal amplification system able to a...

Authors: Unknown author
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Sensitive and specific detection of Aujeszky’s disease virus (ADV), African swine fever virus (ASFV), porcine circovirus type 2 (PCV2) and porcine parvovirus (PPV). This protocol describes the sensitive and specific detection of Aujeszky’s disease virus (ADV), African swine fever virus (ASFV), porcine circovirus type 2 (PCV2) and porcine parvovirus (PPV). The assays target the gD gene (ADV), t...

Authors: Own Document
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Description of how to extract and analyze data generated using the Invader assay. This protocol describes the analysis of invader data generated using either a real-time PCR instrument or an ordinary fluorescent plate reader. As there exist no platform with built in capability to analyze any data generated using an Invader based protocol it has to be extracted and analyzed manually. This can be ba...

Authors: Bernt Hjertner
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Description of how to prepare silica particles for the extraction of nucleic acids. This protocol describes the preparation of Silica particles for the extraction of nucleic acids. DNA and RNA bind efficiently to acid-washed Silica particles (Boom et al., 1990). This procedure rescues the nucleic acids from the main part of debris and interfering agents in serum and organ materials. The current me...

Authors: Thomas Bruun Rasmussen
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This protocol describes extraction of nucleic acids from serum or organ material for subsequent RT-PCR analysis. This protocol describes extraction of nucleic acids from serum or organ material for subsequent RT-PCR analysis. The content of PCR-interfering agents is reduced thus resulting in an increased sensitivity of the PCR. Detection of RNA viruses by PCR is often hampered by the interference ...

Authors: Thomas B Rasmussen
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This protocol describes the detection by one-step RT-PCR (Reverse Transcriptase Polymerase Chain Reaction) with a TaqMan® probe of all the serotypes of AHSV. The primers target a conserved region within the NS1 segment. The PCR product expands 210 bp. The set of primers and the FAM-labelled probes used in this protocol has been designed by Partner 4, UCM, Madrid.

Authors: Belen Rodriguez-Sanchez
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A one-step protocol for detection of of bluetongue virus (BTV) RT-PCR. This protocol describes a one-step protocol for SYBR Green RT-PCR (Reverse Transcriptase Polymerase Chain Reaction) amplification of bluetongue virus (BTV).This new RT-PCR has been developed by Partner 4, UCM, Madrid, for the specific detection of BTV serotype 4. The pair of primers described here target a sequence within the V...

Authors: Belen Rodriguez-Sanchez
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