Pretreatment to avoid positive RT-PCR results with inactivated viruses

J Virol Methods. 2002 Jul;104(2):217-25. doi: 10.1016/s0166-0934(02)00089-7.


Enteric viruses that are important causes of human disease must often be detected by reverse transcription-polymerase chain reaction (RT-PCR), a method that commonly yields positive results with samples that contain only inactivated virus. This study was intended to develop a pretreatment for samples, so that inactivated viruses would not be detected by the RT-PCR procedure. Model viruses were human hepatitis A virus, vaccine poliovirus 1 and feline calicivirus as a surrogate for the Norwalk-like viruses. Each virus was inactivated (from an initial titer of approximately 10(3) PFU/ml) by ultraviolet light, hypochlorite or heating at 72 degrees C. Inactivated viruses, that were treated with proteinase K and ribonuclease for 30 min at 37 degrees C before RT-PCR, gave a negative result, which is to say that no amplicon was detected after the reaction was completed. This antecedent to the RT-PCR method may be applicable to other types of viruses, to viruses inactivated in other ways and to other molecular methods of virus detection.

Publication types

  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Animals
  • Calicivirus, Feline / genetics
  • Calicivirus, Feline / immunology
  • Calicivirus, Feline / isolation & purification*
  • Cats
  • Cell Line
  • Hepatitis A virus / genetics
  • Hepatitis A virus / immunology
  • Hepatitis A virus / isolation & purification*
  • Humans
  • Kidney
  • Macaca mulatta
  • Models, Animal
  • Norovirus / genetics
  • Norovirus / immunology
  • Norovirus / isolation & purification
  • Poliovirus / classification
  • Poliovirus / genetics
  • Poliovirus / immunology
  • Poliovirus / isolation & purification*
  • Polymerase Chain Reaction / methods
  • Reverse Transcriptase Polymerase Chain Reaction*
  • Vaccines, Inactivated
  • Viral Vaccines


  • Vaccines, Inactivated
  • Viral Vaccines