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Dengue is a disease caused by a virus from the family Flaviviradae, carried by a female mosquito of Aedes aegypti species. Dengue fever is widespread in the tropic areas. It caused by rainfall, temperature and unplanned urbanization. According to the ministry of health , almost all provinces in Indonesia are endemic areas of dengue fever. In 2014, up to mid-December Dengue Hemorrhagic Fever (DHF) patients in 34 provinces in Indonesia are 71,668 people and 641. This figure is lower than the previous year, 2013 with 112,511 people and 871 deaths . This disease consists of four types of serotypes, namely DENV-1, DENV-2, DENV-3, and DENV-4. This disease can be identified using a variety of methods, one of the method is Reverse Transcription - Polymerase Chain Reaction (RT-PCR) method. This study aims to determine the ability of Dimethylformamide (DMF), Ethylenediamintetraacetic Acid (EDTA), and Ultrapure H2O as the substitute of Ethanol for precipitation in RNA extraction process. The sample used in this research obtained from Surabaya. RNA extraction itself can be done by using a special kit for RNA extraction. In Reverse Transcription - Polymerase Chain Reaction method, first RNA is extracted and then transcribed back (Reverse Transcription) which then form cDNA that later will be amplified by using PCR method. In this study used specific primers for dengue virus type 3 (DENV-3). The results of this study show that DMF, EDTA, and Ultrapure H2O can be used as the substitute of Ethanol for precipitation on RNA extraction. The result is evidenced by the formation of viral DNA bands on gel electrophoresis results.

Dengue virus distributed in tropical and subtropical regions in the world. DENV viruses are transmitted between humans primarily by Aedes aegypti and Aedes albopictus mosquitoes and are endemic in most areas in which the vectors occur. Four serotypes of dengue virus are DENV-1, DENV-2, DENV-3 and DENV-4. DENV-2 is comprised of six genotypes. Ethanol precipitation is a commonly used technique for concentrating and de-salting nucleic acids (DNA or RNA) preparations in aqueous solution. RNA isolation by combining Guanidinium thiocyanate and phenol reported has been reported. In this report, we investigated RNA isolation from DENV-2 using QIAamp Mini Kit with 2-Isopropanol, Methanol, Chloroform precipitation solvent. Electrophoregram showed DNA band as the result of RNA isolation with methanol and 2-isopropanol are produced quite well. Dna band of the of RNA isolation with chloroform solvent has the lowest intensity than methanol and 2-isopropanol. This study showed that methanol and 2-isopropanol can used as precipitation solvent for isolating RNA.