CLONING AND SEQUENCING OF PFLDH GENE FOR MALARIA DIAGNOSIS AND APPLICATION OF PROTISTA MATERIAL IN HIGH SCHOOL

• Main Author: Siti Hotimah, Yeni
• Format: Theses
• Language: Indonesia
• Subjects: Kimia
• Online Access: https://digilib.itb.ac.id/gdl/view/36265
• Institution: Institut Teknologi Bandung

Plasmodium is a genus of Sporozoa phylum in the Protist kingdom. Plasmodium topic is introduced at high school in the tenth grade of science classes, where Plasmodium life cycle as a parasite that causes malaria is discussed. Malaria is one of the most important issues in global health problems that can cause morbidity and mortality in various countries in the world including Indonesia, especially eastern Indonesia such as Jayapura, which is an endemic area for malaria. Fast and accurate diagnosis is the basis for malaria prevention. The tool currently being developed is the Rapid Diagnostic Test (RDT) based on immunochromatography. One of the antigens marker used for diagnosis is Lactate dehydrogenase (LDH) because it is produced in large quantities both in the sexual and asexual stages on the glycolytic pathway in the parasite. This research aims to introduce the students to the process of diagnosing malaria with an immunochromatographic based RDT through animation that can be used as a learning media for Protista topic. Animation video has been successfully created and can be seen on https://youtu.be/mr4CIBGHM-M. To evaluate whether animated video could help the student to understand the topic easier, tests were conducted on two groups of students from high school in Bandung. The two groups were given the same question, the first group was given an explanation in an animated video while the other group was given an explanation in a text. Results from the two test groups were compared. The evaluation results show that animated video can improves the average scores, more informative, more interesting and easier to understand than information in a text. In this study, we also performed cloning and sequencing of the Plasmodium falciparum lactate dehydrogenase (PfLDH) gene, which were carried out from a blood sampel of a Plasmodium falciparum-infected-patient from Jayapura. Recombinant pGEM-T-PfLDH has been successfully constructed. PfLDH gene was amplified using PCR method, then was ligated to vector cloning pGEM-T. The pGEM-T-PfLDH plasmid then was transformed into E. coli TOP10F’ using kejut panas method, and confirmed by colony PCR as well as restriction analysis. Sequencing analysis of pGEM-T-PfLDH showed 100% similarity with PF3D7_1324900.