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.
Mol Biol Rep. 2011 Nov;38(8):4979-83. doi: 10.1007/s11033-010-0642-9. Epub 2010 Dec 15.
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.
从人血浆中纯化登革病毒 RNA 可用于研究和临床目的。登革热在巴西的圣埃斯皮里图州流行，且正逐渐成为一个难以控制的公共卫生问题。巴西目前存在登革病毒 1、2 和 3 型，最近报告称登革病毒 4 型已进入巴西边境。该病毒在疫情爆发期间迅速传播，每年有数千人感染，由于出血性登革热，死亡人数被低估。由于这种疾病主要影响发展中国家，因此必须找到一种强大、快速且低成本的病毒核酸纯化方法。在本文中，我们比较了两种从临床诊断为登革热的患者的血清/血浆中提取 RNA 的方法。使用 QIAamp(®) UltraSens Virus Kit（Qiagen Inc.，美国瓦伦西亚）和较便宜的 Chomczynski-Sacchi 法分析了总共 47 个样本。核酸纯化后，用登革病毒 2 型特异性引物进行逆转录和聚合酶链反应扩增。在我们的地理位置，这种亚型最为普遍。用 Chomczynski-Sacchi 技术提取 RNA 时，有 34 个样本呈阳性，而用 QIAamp(®) UltraSens Virus Kit 时，只有 27 个样本呈阳性。这些结果有利于使用更经济实惠的技术来纯化病毒 RNA，这对于发展中国家尤为重要。
