DETEKSI DAN IDENTIFIKASI DENV-1 MENGGUNAKAN REVERSE TRANSCRIPTASE POLYMERASE CHAIN REACTION: SEBUAH STUDI VIROLOGI

Abstract

Dengue virus (DENV) as an etiology of Dengue Fever/Dengue Hemorrhagic Fever, is divided into four serotypes: DENV-1, DENV-2, DENV-3, and DENV-4, which all of them could be found in Indonesia. The research was done to measure the frequency of DENV-1 infection. The research had never been done before, thus it has a benefit as data base about this virus. DENV extraction process was taken from human plasma of 100 patients whom contracted Dengue Fever/Dengue Hemorrhagic Fever, where the number of samples was determined by proportional formula. The research took places from three hospitals in Medan, and then the product underwent a serotype identification process using Reverse Transcriptase Polymerase Chain Reaction (RT-PCR), using cross sectional design the result would be archived and presented in percentage data. There are 2% positive of serotype 1 of 100 samples in 5 to 9 years old age class and 15 to 44 years old age class, respectively and the gender are males. This result was found post extraction of the virus’ Ribonucleic Acid (RNA) from infected cells. The extraction process was initialized by preparing Qiagen protease by using QIAamp® Viral RNA Mini Kit from Qiagen®. Next, the human plasma was inserted into eppendorf tubes, in order to form a supernatant by centrifugation. The supernatant and Qiagen® protease underwent incubation processes and centrifugations and the final product was stored in -70^oC temperature. Reverse transcriptase procedure was engaged after to get complement Deoxyribonucleic Acid (cDNA), and then got amplified with PCR procedure and the final product was stored in 4^oC temperature. Finally, it underwent electrophoresis process and visualized with gel imaging instrument. It was concluded, of 4 serotypes, DENV-1 percentage is low. However, the awareness of dengue infection has to be kept high, considering its possibility in developing massive bleeding complication caused by secondary heterologous infection due to another serotype infection.