SYNTHESIS OF RECOMBINANT PROTEIN NS1 AND EDIII GOLD CONJUGATE FOR DEVELOPMENT DENGUE IGG/IGM DIAGNOSTIK TEST
Around 50-100 million Dengue Hemorrhagic Fever (DHF) infections occur in more than 100 endemic countries. For the period January - October 2019, there were 110,921 dengue cases occurring in Indonesia. Indonesia is one of the countries categorized as having very high dengue cases, so an effort is nee...
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| Format: | Theses |
| Language: | Indonesia |
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| Online Access: | https://digilib.itb.ac.id/gdl/view/47724 |
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| Institution: | Institut Teknologi Bandung |
| Language: | Indonesia |
| Summary: | Around 50-100 million Dengue Hemorrhagic Fever (DHF) infections occur in more than 100 endemic countries. For the period January - October 2019, there were 110,921 dengue cases occurring in Indonesia. Indonesia is one of the countries categorized as having very high dengue cases, so an effort is needed to control dengue virus infection by detecting dengue infection early and followed by proper treatment. One specific, sensitive, safe, and inexpensive laboratory diagnostik method is a diagnostik kit that can detect the presence of dengue antibodies (IgM and IgG). One very important component to produce this diagnostik kit is conjugate of gold nanoparticles with proteins. A gold nanoparticle of about 40 nm in red colour has functions as a coloring agent so that positive results can be monitored as a purplish red line on the membran kit. This gold conjugate is made by conjugating gold nanoparticles with a protein that acts to recognize IgM or IgG antibodies in the patient's serum. The recombinant proteins NS1 and EDIII were chosen because they have quite high antigenic properties. The DENV gold conjugate was obtained from the conjugation of 2 ?g of NS1 and EDIII proteins per ml of colloidal gold. Characterization of the gold conjugate using a UV-VIS spectrophotometer showed that there was a shift in the maximum wavelength from 525 nm to around 530 nm due to conjugation, which indicated the formation of complexes with gold nanoparticles. Based on measurement of particle diameter / size using Dynamic Light Scateering (DLS), there is an increase in the size of gold nanoparticles from 43 nm to 79 nm for the gold-NS1 conjugate and 68 nm for the gold-EDIII conjugate.
In addition, the characterization by the Dot Blot show the ability of the gold conjugate to recognize the serum of positive Dengue patients. |
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