Tesis

Produksi dan Karakterisasi Pseudovirus Nipah Sebagai Pengembangan Uji Netralisasi = Production and Characterization of Nipah Pseudovirus as A Neutralization Test Development.

Virus Nipah (NiV) merupakan paramyxovirus yang menyebabkan penyakit pernapasan akut dan ensefalitis parah pada spektrum luas vertebrata, termasuk manusia. Karena tingkat patogenisitas dan kematiannya yang tinggi, NiV dianggap sebagai prioritas kesehatan global oleh Organisasi Kesehatan Dunia (WHO), sehingga memerlukan penanganan di fasilitas penahanan Biosafety Level-4 (BSL-4), dan membutuhkan waktu beberapa hari untuk menyelesaikannya. Saat ini belum tersedia vaksin atau agen terapeutik yang efektif. Penelitian ini dilakukan untuk membuat pseudovirus NiV vektor tulang punggung lentivirus HIV dan di fasilitas tingkat keamanan hayati 2. Plasmid ekspresi rekombinan protein NiV G dan protein NiV F, psPAX2, dan pLenti CMV Puro LUC (w168-1) ditransfeksi secara transien ke dalam sel HEK 293T selama 72 jam untuk menghasilkan pseudovirus NiV. Pseudovirus NiV dibuat menggunakan sel HEK 293T. Spesifisitas ekspresi protein NiV G dan NiV F ditunjukkan dengan indirect immunofluorescence dan western blot. Partikel pseudovirus menunjukkan morfologi yang jelas di bawah scanning electron microscope (SEM) dan memiliki intensitas luciferase 2.8 × 106 RLU. Pseudovirus NiV berhasil dibuat menggunakan sistem vektor HIV dan hasil penelitian ini diharapkan dapat memvalidasi keamanan dan spesifisitas tes netralisasi terhadap pseudovirus NiV.
Kata kunci: Virus Nipah, Pseudovirus, Protein G, Protein F


Nipah virus (NiV) is a paramyxovirus that causes acute respiratory disease and severe encephalitis in a broad spectrum of vertebrates, including humans. Due to its high pathogenicity and mortality, NiV is considered a global health priority byt he World Health Organization (WHO), requiring treatment in a Biosafety Level-4 (BSL-4) containment facility, and taking several days to complete. There is currently No. effective vaccine or therapeutic agent available. This study was conducted to create the HIV lentivirus backbone vector NiV pseudovirus and in a biosafety level 2 facility. Recombinant expression plasmids of NiV G protein and NiV F protein, psPAX2, and pLenti CMV Puro LUC (w168-1) were transiently transfected into HEK 293T cells for 72 hours to generate NiV pseudovirus. NiV pseudoviruses were generated using HEK 293T cells. The specificity of NiV G and NiV F protein expression was demonstrated by indirect immunofluorescence and western blot. The pseudovirus particles showed clear morphology under scanning electron microscope (SEM) and had a luciferase intensity of 2.8 × 106 RLU. NiV pseudovirus was successfully generated using HIV vector system and the results of this study are expected to validate the safety and specificity of the neutralization test against NiV pseudovirus.
Keywords: Nipah virus, Pseudovirus, G Protein, F Protein