PENGEMBANGAN METODE LOOP-MEDIATED ISOTHERMAL AMPLIFICATION OF DNA UNTUK DETEKSI Megalocytivirus TERHADAP GEN MAJOR CAPSID PROTEIN PADA KERAPU
| Main Authors: | , Sumino, , Dr. Ir. Murwantoko, M.Si |
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| Format: | Thesis NonPeerReviewed |
| Terbitan: |
[Yogyakarta] : Universitas Gadjah Mada
, 2014
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| Subjects: | |
| Online Access: |
https://repository.ugm.ac.id/133318/ http://etd.ugm.ac.id/index.php?mod=penelitian_detail&sub=PenelitianDetail&act=view&typ=html&buku_id=73911 |
Daftar Isi:
- Grouper fish is one of the most cultivated fishery commodity in Indonesia due to its high economical value. Infectious disease caused by Megalocytivirus is one of the bottle neck in aquaculture. Thus, quick, simple and cheap viral detection method is necessary. Loop-Mediated Isothermal Amplification (LAMP) of DNA is developed to detect Megalocytivirus isolate of Indonesia with Major Capsid Protein (MCP) gene as the target amplification using strand displacement of activity at isothermal condition of DNA polymerase. The optimization condition of LAMP reaction includes temperature and betain concentration. The optimization studied were applied for specificity, sensitivity, and sample tests. Grouper fish samples were collected from Batam, Lampung, Jepara, and Bali. Sensitivity result of LAMP was compared with PCR method. The samples were also being tested using PCR and histopathology to confirm the result. The results of the study showed that the optimum condition for LAMP reaction of Megalocytivirus isolate of Indonesia is on 62,50 C and betain concentration of 1 M. LAMP is highly specific and its sensitivity is ten times higher than PCR on detecting Megalocytivirus samples. Detection of Megalocytivirus on grouper fishes collected from Batam, Lampung, Jepara, and Bali using PCR and LAMP provided similar results. Among 22 samples tested, 8 were positive for Megalocytivirus although based on histopahology examination, the samples did not show any pathognomonic of Megalocytivirus infection.