JoVE Journal

Immunology and Infection

This content is Open Access.

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

28.2K Views

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09:57 min

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March 17th, 2016

DOI :

10.3791/53672-v

March 17th, 2016

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Erika S. Hue¹^,²^,³, Christine I. Fortier¹^,³, Aurélie M. Laurent¹, Yann F. Quesnelle¹, Guillaume D. Fortier¹^,²^,³, Loïc J. Legrand¹^,²^,³, Stephane L. Pronost¹^,²^,³

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

Summary

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.

Explore More Videos

Equine Herpesvirus 2

Respiratory Fluids

Nucleic Acid Extraction

Diagnostic Method

Quantitative Data

Respiratory Disease

Chapters in this video

0:05

Title

0:55

Extraction of Nucleic Acids

2:03

Amplification Procedure

3:57

Limit of Detection of qRT-PCR

5:54

Limit of Detection of the Method Determined by Two Independent Trials

7:04

Results: Validation of Quantitative PCR Method for Equid Herpesvirus-2 Diagnosis

8:45

Conclusion

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