Pretreatment to avoid positive RT-PCR results with inactivated viruses.
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.
Nuanualsuwan
Suphachai
S
Department of Population Health and Reproduction, School of Veterinary Medicine, University of California, 1 Shields Avenue, Davis, CA 95616-8743, USA.
Cliver
Dean O
DO
D000818:Animals; D017927:Calicivirus, Feline / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D002415:Cats; D002460:Cell Line; D030041:Hepatitis A virus / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D006801:Humans; D007668:Kidney; D008253:Macaca mulatta; D023421:Models, Animal; D029322:Norovirus / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification; D017955:Poliovirus / Q000145:classification / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction / Q000379:methods; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015164:Vaccines, Inactivated; D014765:Viral Vaccines
D015164:Vaccines, Inactivated; D014765:Viral Vaccines