RISZA HARTAWAN
Indonesia Research Center for Veterinary Science Indonesian Agency for Agricultural Research and Development Jalan R.E. Martadinata No. 30, PO Bx 52 Bogor 16114, West Java, Indonesia

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IN SILICO ANALYSIS OF SMALL INTERFERING RNA TARGETING THE NUCLEOPROTEIN GENE OF INFLUENZA VIRUSES Hartawan, Risza; Ratnawati, Atik; Sendow, Indrawati; Dharmayanti, Ni Luh Putu Indi
Jurnal Bioteknologi & Biosains Indonesia (JBBI) Vol. 11 No. 2 (2024)
Publisher : BRIN - Badan Riset dan Inovasi Nasional

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.55981/jbbi.2024.8521

Abstract

Small interfering RNA (siRNA) is a promising therapeutic against viral infection, includ-ing Influenza viruses. However, the Influenza viruses have massive variants with high mutation rates. Therefore, the siRNAs could be futile against newly emerging viruses. Thus, this study aimed to analyze siRNAs targeting the nucleoprotein gene of Influen-za viruses. Using bioinformatic analyses, the siRNAs were simulated against 5 sub-types of Influenza viruses, including H1N1, H3N2, H5N1, H7N9, and H9N2. Bioinfor-matic tools for the folding structure of messenger RNA were utilized to select effective siRNA. As a result, 32 siRNA sequences targeting the nucleoprotein gene were identi-fied. The precision medicine concept seems applied to the siRNA treatment for the In-fluenza virus since each siRNA is effective in its respective virus target. Based on the nucleotide mismatch parameter, most siRNA does not have coverage for the multiple infections of all five subtypes of Influenza viruses, except for NP1089 and NP1496. Later, the secondary and tertiary structure analysis of messenger RNA demonstrated that siRNA has different circumstances in its RNA target position. Therefore, siRNA mapping based on the RNA folding structure approach provides a tool for selecting more effective sequences against Influenza virus infection. Both siRNA NP1089 and NP1496 were predicted to have similar effectivity in knocking down Influenza virus in-fection. Moreover, the cocktail application of siRNA treatment may be effective as an alternative strategy in matching co-infection of multiple Influenza virus subtypes.
Karakterisasi Kultur Virus African Swine Fever Sampel Lapang Indonesia Menggunakan Kultur Sel Primer Leukosit Babi Ratnawati, Atik; Hartawan, Risza; Sendow, Indrawati; Assadah, Nur Syabiq; Nuradji, Harimurti; Dharmayanti, Ni Luh Putu Indi; Wibawan, I Wayan Teguh; Mayasari, Ni luh Putu Ika
Jurnal Sain Veteriner Vol 43, No 3 (2025): Desember
Publisher : Faculty of Veterinary Medicine, Universitas Gadjah Mada bekerjasama dengan PB PDHI

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22146/jsv.109976

Abstract

African Swine FeverĀ (ASF) merupakan penyakit viral yang sangat menular dan mematikan pada babi, yang disebabkan oleh virus African Swine Fever (ASF). Sejak tahun 2018, penyakit ini telah menyebar dan menyebabkan konsekuensi sosial ekonomi yang besar terhadap industri babi di beberapa negara Asia, termasuk China, Vietnam, dan Indonesia. Penelitian ini bertujuan untuk melakukan isolasi virus ASF dari sampel lapang menggunakan kultur sel primer leukosit babi, identifikasi karakteristik hasil propagasi virus ASF secara in vitro dan respon sel leukosit terhadap inokulasi virus ASF. Sel leukosit dikoleksi dari darah babi donor yang sehat dan dikultur dalam medium plasma homolog. Propagasi dilakukan dengan menginokulasikan sampel lapang dengan kode Indonesia/2022/Pig/PSJ ke kultur sel primer leukosit babi yang konfluen (70-80%). Pengamatan morfologi sel dilakukan dengan menggunakan mikroskop cahaya setiap 24, 48, 72, 96,120, 144, and 164 jam pasca inokulasi (jpi). Sampel lapang dari kultur sel primer leukosit babi dipurifikasi dengan menggunakan metode Percoll. Pelet virus di deteksi virus ASF dengan menggunakan uji qPCR. Hasil penelitian menunjukkan adanya perubahan morfologis pada sel primer leukosit yang terinfeksi, dengan adanya reaksi hemadsorpsi (HAD) yang teramati pada 48 jpi, dibandingkan dengan sel kontrol yang tidak terinfeksi. Pengikatan eritrosit babi ke permukaan sel yang terinfeksi virus ASF, membentuk rosette-like structure. Reaksi hemadsorpsi (HAD) dapat diamati setelah 2 kali blind passage. Purifikasi virus ASF menggunakan Percoll dapat meningkatkan kemurnian virus yang ditandai dengan nilai Ct yang lebih rendah dibandingkan dengan supernatant hasil kultur sel primer leukosit babi.