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Development and Validation of a Quantitative PCR Method for Equid Herpesvirus-2 Diagnostics in Respiratory Fluids
JoVE Journal
Immunology and Infection
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JoVE Journal Immunology and Infection
Development and Validation of a Quantitative PCR Method for Equid Herpesvirus-2 Diagnostics in Respiratory Fluids

Development and Validation of a Quantitative PCR Method for Equid Herpesvirus-2 Diagnostics in Respiratory Fluids

DOI:
10.3791/53672-v

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09:57 min

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March 17, 2016

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, , , , , ,
1LABÉO Frank Duncombe, 2Unité de Risques Microbiens (U2RM, EA 4655),Normandy University, 3Hippolia Foundation

Chapters

  • 00:05Title
  • 00:55Extraction of Nucleic Acids
  • 02:03Amplification Procedure
  • 03:57Limit of Detection of qRT-PCR
  • 05:54Limit of Detection of the Method Determined by Two Independent Trials
  • 07:04Results: Validation of Quantitative PCR Method for Equid Herpesvirus-2 Diagnosis
  • 08:45Conclusion

Summary

Automatic Translation

Automatic Translation

Here, we present a protocol for the development and validation of a quantitative PCR method used for the detection and quantification of EHV-2 DNA in equine respiratory fluids. The EHV-2 qRT-PCR validation protocol involves a three-part procedure: development, characterization of qRT-PCR assay alone, and characterization of the whole analytical method.

Tags

QPCR MethodEquid Herpesvirus-2 DiagnosticsViral LoadRespiratory FluidsDiagnostic FieldsQuantitative DataInterpretative AidsSensitive MethodSpecific MethodRapid MethodAFNOR Norme NF U47-600Erika HueYoung ResearcherNucleic Acids ExtractionLysis SolutionEthanolSilica ColumnCentrifugeWash Buffers AW1 And AW2Elution Buffer

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