PENGEMBANGAN UJI DIAGNOSTIK MOLEKULER VIRUS DENGUE BERBASIS SYBR GREEN REAL-TIME PCR UNTUK DIAGNOSIS AWAL DEMAM BERDARAH

• Main Author: Khoirunnisa, Karimatu
• Format: Final Project
• Language: Indonesia
• Online Access: https://digilib.itb.ac.id/gdl/view/48705
• Institution: Institut Teknologi Bandung

Incidence rate of dengue fever in Indonesia is high and shows increasing trend year to year. The severity of this fever can be decreased by early diagnostic test. Unfortunately, Indonesia has not had the kit for early diagnostic for dengue fever. So, this research aims to develop diagnostic test using Real-Time PCR in which has good specificity and easy to be used. Dengue fever caused by the infection of dengue virus, a virus from Flaviviridae group that has positive ssRNA genome. The vector of this virus is Aedes aegypti and Aedes albopictus. In this research amplification target region designed by multiple sequence alignment. Primer designing done in silico by using Bioedit software. Positive control designed by constructing amplification target in backbone plasmid pUC57. Amplification preliminary test done by PCR method. Confirmation test of primer designed done with Indonesia-Taiwan collaboration positive dengue fever RNA sample of the ....... 10542-10683 nucleotide sequence as the amplification target region candidate of Primer set 1 and Primer set 1 as primer pair candidate. PCR test showed amplicon band with size 142 bp. Real-time PCR confirmation test showed amplification curve of the sample used with melting temperature around 85OC. In conclusion this research successfully developed diagnostic test for early diagnosis of dengue fever. However, linearity test should be done and the detection limit of the diagnostic test developed should be determined.