title: An optimized method for elution of enteroviral RNA from a cellulose-based substrate.
abstract: The Flinders Technology Australia (FTA) Elute Card is a commercial product that facilitates the collection, transport, archiving and processing of nucleic acids from a wide variety of biological samples at room temperature. While the cards have been designed so that sterile/deionized water can elute DNA easily, they are not suitable for some less stable RNAs. This study was undertaken to determine the optimal conditions such as the buffer type, buffer pH and incubation temperature for the elution of enteroviral RNA from FTA Elute Cards prior to quantitative analysis using real-time PCR (qPCR) or consensus degenerate hybrid oligonucleotide primer VP1 RT-semi nested PCR (CODEHOP VP1 RT-snPCR). TE-1 (pH 8.0), rather than sterile water, was the best buffer for high efficiency elution of enteroviral RNA at 95°C. However, as the estimated recovery rate of viral RNA eluted from the cards averaged to be only 6.1%, enterovirus assays using FTA elution should be considered qualitative, especially at low virus titers, and therefore the results of the assay should be interpreted carefully.
authors.affiliation: Shandong Provincial Key Laboratory of Infectious Diseases Control and Prevention, Shandong Center for Disease Control and Prevention, 16992 Jingshi Road, Jinan 250014, China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Li
authors.forename: Hiromu
authors.initials: H
authors.lastname: Yoshida
authors.forename: Lu
authors.initials: L
authors.lastname: Wang
authors.forename: Zexin
authors.initials: Z
authors.lastname: Tao
authors.forename: Haiyan
authors.initials: H
authors.lastname: Wang
authors.forename: Xiaojuan
authors.initials: X
authors.lastname: Lin
authors.forename: Aiqiang
authors.initials: A
authors.lastname: Xu
mesh_terms: D002021:Buffers; D002482:Cellulose / Q000378:metabolism*; D004770:Enterovirus / Q000235:genetics / Q000302:isolation & purification*; D004769:Enterovirus Infections / Q000175:diagnosis / Q000821:virology; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008967:Molecular Biology / Q000379:methods*; D012367:RNA, Viral / Q000302:isolation & purification* / Q000378:metabolism*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods; D013048:Specimen Handling / Q000379:methods*; D013696:Temperature
chemical_list: D002021:Buffers; D012367:RNA, Viral; D002482:Cellulose