[Validation of a real time RT-PCR assay to detect foot-and-mouth disease virus and assessment of its performance in acute infection]

Rev Argent Microbiol. 2014 Jul-Sep;46(3):188-95. doi: 10.1016/S0325-7541(14)70071-9. Epub 2014 Oct 15.
[Article in Spanish]

Abstract

A specific real time reverse transcription polymerase chain reaction (RT-PCRrt) for the detection of foot-and-mouth disease virus was validated using the LightCycler thermocycler 2.0 and its reagents as recommended by the World Organization for Animal Health and was assessed for the detection of the virus in acute infection of cattle experimentally vaccinated and challenged with virus A Argentina/2001 or A24 Cruzeiro. The technique proved to be robust, showing coefficients of variation lower than 4% for different ARN extractions, days or repetitions and was able to detect up to 0,4 TCID 50%, and/or up to 100 RNA molecules. In probang samples, diagnostic sensitivity was 93.1 (95% CI 86.5-96.6) and diagnostic specificity 100 (95% CI 96.3-100). The results of the challenge in vaccinated or multivaccinated bovines showed that although there were high levels of clinical protection in the vaccinated group, FMDV could be detected in all challenged groups. However, detection was 100 times lower in immunized animals.

Keywords: Diagnosis; Diagnóstico; Fiebre Aftosa; Foot-and-mouth disease; RT-PCRtr; Real time RT-PCR.

Publication types

  • Research Support, Non-U.S. Gov't
  • Validation Study

MeSH terms

  • Acute Disease
  • Animals
  • Body Fluids / virology
  • Cattle
  • Cattle Diseases / diagnosis*
  • Cattle Diseases / virology
  • Esophagus / virology
  • Foot-and-Mouth Disease / diagnosis*
  • Foot-and-Mouth Disease / virology
  • Foot-and-Mouth Disease Virus / classification
  • Foot-and-Mouth Disease Virus / genetics
  • Foot-and-Mouth Disease Virus / immunology
  • Foot-and-Mouth Disease Virus / isolation & purification*
  • Pharynx / virology
  • RNA, Viral / analysis
  • Real-Time Polymerase Chain Reaction / instrumentation
  • Real-Time Polymerase Chain Reaction / methods*
  • Reproducibility of Results
  • Reverse Transcriptase Polymerase Chain Reaction / instrumentation
  • Reverse Transcriptase Polymerase Chain Reaction / methods*
  • Sensitivity and Specificity
  • Specimen Handling
  • Vaccination / veterinary
  • Viral Vaccines / immunology

Substances

  • RNA, Viral
  • Viral Vaccines