title: Discrimination of infectious hepatitis A viruses by propidium monoazide real-time RT-PCR.
abstract: The discrimination of infectious and inactivated viruses remains a key obstacle when using quantitative RT-PCR (RT-qPCR) to quantify enteric viruses. In this study, propidium monoazide (PMA) and RNase pretreatments were evaluated for the detection and quantification of infectious hepatitis A virus (HAV). For thermally inactivated HAV, PMA treatment was more effective than RNase treatment for differentiating infectious and inactivated viruses, with HAV titers reduced by more than 2.4 log(10) units. Results showed that combining 50 μM of PMA and RT-qPCR selectively quantify infectious HAV in media suspensions. Therefore, PMA treatment previous to RT-qPCR detection is a promising alternative to assess HAV infectivity.
authors: Departamento de Biotecnología, Instituto de Agroquímica y Tecnología de Alimentos, Avda. Agustín Escardino, 7. Paterna, Valencia, Spain. gloriasanchez@iata.csic.es
authors: Gloria
authors: G
authors: Sánchez
authors: Patricia
authors: P
authors: Elizaquível
authors: Rosa
authors: R
authors: Aznar
mesh_terms: D000818:Animals; D001386:Azides / Q000494:pharmacology*; D002460:Cell Line; D006506:Hepatitis A / Q000821:virology*; D030041:Hepatitis A virus / Q000187:drug effects / Q000235:genetics / Q000302:isolation & purification* / Q000472:pathogenicity; D006358:Hot Temperature; D006801:Humans; D006997:Hypochlorous Acid / Q000494:pharmacology; D008253:Macaca mulatta; D011419:Propidium / Q000031:analogs & derivatives* / Q000494:pharmacology; D012367:RNA, Viral / Q000032:analysis / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012260:Ribonucleases / Q000378:metabolism*; D038301:Virus Inactivation
chemical_list: D001386:Azides; D012367:RNA, Viral; C533957:propidium monoazide; D011419:Propidium; D006997:Hypochlorous Acid; D012260:Ribonucleases