title: Dengue virus RNA purification from human plasma: a comparison of two techniques.
abstract: Dengue virus RNA purification from human plasma is useful for research and clinical purposes. Dengue is endemic in the Espirito Santo State, Brazil, and it is progressively becoming a hard-to-control public health problem. Dengue virus types 1, 2 and 3 are currently found in Brazilian territory, and recently Dengue virus type 4 has been reported to enter Brazilian borders. This virus spreads rapidly during epidemic outbreaks, and thousands of patients are infected annually, with an underestimated number of deaths in consequence of hemorrhagic Dengue. Because this disease affects mainly developing countries, it is imperative that a robust, rapid and low cost method for viral nucleic acid purification is found. In this manuscript we compare two RNA extraction methods from serum/plasma of patients with clinical diagnosis of dengue. The QIAamp(®) UltraSens Virus Kit (Qiagen Inc., Valencia, USA) and the less expensive Chomczynski-Sacchi method were used to analyze a total of 47 samples. After nucleic acid purification, reverse transcription and polymerase chain reaction amplification with dengue virus type 2 specific primers were performed. This subtype is the most prevalent in our geographical location. Thirty-four samples were positive when RNA was extracted by the Chomczynski-Sacchi technique, whereas only 27 of these were positive when the QIAamp(®) UltraSens Virus Kit was used. These results favor the utilization of the more affordable technique for the purification of viral RNA, which is especially important for developing countries.
authors.affiliation: Núcleo de Genética Humana e Molecular, Departamento de Ciências Biológicas, Centro de Ciências Humanas e Naturais, Universidade Federal do Espírito Santo, Av. Marechal Campos, 1468 - Campus de Maruípe, Vitória, ES 29040-090, Brazil.
authors.forename: Raquel Spinassé
authors.initials: RS
authors.lastname: Dettogni
authors.forename: Iúri Drumond
authors.initials: ID
authors.lastname: Louro
mesh_terms: D001483:Base Sequence; D003716:Dengue Virus / Q000235:genetics*; D006801:Humans; D008967:Molecular Biology / Q000379:methods*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D012367:RNA, Viral / Q000097:blood* / Q000235:genetics / Q000302:isolation & purification*; D017422:Sequence Analysis, DNA
chemical_list: D012367:RNA, Viral