Karakterisasi Kultur Virus African Swine Fever Sampel Lapang Indonesia Menggunakan Kultur Sel Primer Leukosit Babi

Atik Ratnawati; Risza Hartawan; Indrawati Sendow; Nur Syabiq Assadah; Harimurti Nuradji; Ni Luh Putu Indi Dharmayanti; I Wayan Teguh Wibawan; Ni luh Putu Ika Mayasari

doi:10.22146/jsv.109976

Karakterisasi Kultur Virus African Swine Fever Sampel Lapang Indonesia Menggunakan Kultur Sel Primer Leukosit Babi

https://doi.org/10.22146/jsv.109976

Atik Ratnawati⁽¹⁾, Risza Hartawan⁽²⁾, Indrawati Sendow⁽³⁾, Nur Syabiq Assadah⁽⁴⁾, Harimurti Nuradji⁽⁵⁾, Ni Luh Putu Indi Dharmayanti⁽⁶⁾, I Wayan Teguh Wibawan⁽⁷⁾, Ni luh Putu Ika Mayasari^(8*)

(1) Sekolah Kedokteran Hewan dan Biomedis, IPB University
(2) Badan Riset dan Inovasi Nasional
(3) Badan Riset dan Inovasi Nasional
(4) Kementerian Pertanian
(5) Badan Riset dan Inovasi Nasional
(6) Badan Riset dan Inovasi Nasional
(7) Sekolah Kedokteran Hewan dan Biomedis, IPB University
(8) Sekolah Kedokteran Hewan dan Biomedis, IPB University
(*) Corresponding Author

Abstract

African Swine Fever (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.

Keywords

Blind passage; hemadsorpsi; purifikasi Percoll; qPCR; virus ASF

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DOI: https://doi.org/10.22146/jsv.109976