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Inhibitors of RT-PCR in serum.

D S Konet1, J M Mezencio, G Babcock

  • 1Plum Island Animal Disease Center, Greenport, NY 11944, USA.

Journal of Virological Methods
|January 22, 2000
PubMed
Summary

Substances in animal sera inhibit reverse transcription polymerase chain reaction (RT-PCR) amplification of foot-and-mouth disease virus RNA. This interference stems from direct interaction with the viral RNA, not the RT-PCR enzymes.

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Area of Science:

  • Veterinary Virology
  • Molecular Biology
  • Biochemistry

Background:

  • Foot-and-mouth disease virus (FMDV) poses a significant threat to livestock globally.
  • Accurate and rapid detection of FMDV is crucial for disease control.
  • Reverse transcription polymerase chain reaction (RT-PCR) is a common method for viral RNA detection.

Purpose of the Study:

  • To investigate the cause of inhibition observed during RT-PCR amplification of FMDV RNA.
  • To identify whether serum components or RT-PCR reagents are responsible for the observed inhibition.

Main Methods:

  • RT-PCR amplification of FMDV RNA.
  • Analysis of serum components from various animal species.
  • Assays to differentiate between RNA-target and enzyme-target inhibition.

Main Results:

  • Substances present in sera from multiple species significantly inhibited FMDV RNA amplification via RT-PCR.
  • Evidence suggests direct interaction between serum substances and viral RNA.
  • No significant inhibition was observed towards the enzymes utilized in the RT-PCR process.

Conclusions:

  • Serum-derived factors can interfere with FMDV RNA detection by RT-PCR.
  • The inhibitory mechanism involves direct binding to viral RNA, impacting amplification efficiency.
  • Understanding these inhibitors is important for optimizing diagnostic protocols for FMDV.

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