title: Establishing conditions for the storage and elution of rabies virus RNA using FTA(®) cards.
abstract: The Flinders Technology Associates filter paper cards (FTA(®) cards) can be used to store nucleic acid from various samples and are easily portable. However, RNA is physicochemically unstable compared with DNA, and appropriate methods have not been established for storage and extraction of RNA from FTA(®) cards. The present study investigated the optimum conditions for storage and elution of viral RNA (vRNA) using rabies virus (RABV) applied to FTA(®) cards. When TE buffer was used, the elution rates of vRNA increased with the length of the elution time. When the cards were stored at -80 °C or -20 °C, vRNA was stable over 3 months. Degradation of vRNAs occurred following storage at 4 °C and room temperature, suggesting that RNA should be extracted from cards as soon as possible if no freezer is available. When we tried to amplify vRNA from RABV-infected animal brains applied to FTA(®) cards and stored at -80 °C for 6 months, we did not detect any amplified products with the primer set for 964 bp of RABV N gene. However, we were able to detect amplified products by increasing the elution time of vRNA from FTA(®) cards from 30 min to 24 hr or by changing the primer sets to amplify 290 bp of N gene. Thus, we recommend extending the elution time for damaged or low concentration samples in FTA(®) cards.
authors: Nihon University Veterinary Research Center, 1866 Kameino, Fujisawa, Kanagawa 252-0880, Japan.
authors: Takeo
authors: T
authors: Sakai
authors: Ayako
authors: A
authors: Ishii
authors: Takao
authors: T
authors: Segawa
authors: Yukihiko
authors: Y
authors: Takagi
authors: Yuki
authors: Y
authors: Kobayashi
authors: Takuya
authors: T
authors: Itou
mesh_terms: D000818:Animals; D001921:Brain; D004655:Emulsions; D051379:Mice; D010209:Paper; D012367:RNA, Viral / Q000737:chemistry* / Q000378:metabolism; D011820:Rabies virus / Q000302:isolation & purification* / Q000378:metabolism; D013048:Specimen Handling / Q000379:methods*; D013696:Temperature
chemical_list: D004655:Emulsions; D012367:RNA, Viral
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