Pertumbuhan Virus Newcastle Disease Isolat Lokal di Kultur Sel LestarI Baby Hamster Kidney-21 (BHK-21)

Abstract

Virus Newcastle Disease merupakan penyebab penyakit patogen pada unggas, namun di balik sifat patogennya terhadap unggas, virus ND memiliki efek onkolitik yang dapat dimanfaatkan sebagai agen viroterapi. Penelitian ini bertujuan menguji kemampuan pertumbuhan virus ND genotipe VII asal Sulawesi Selatan tahun 2019 secara in-vitro dalam sel lestari BHK-21. Sel BHK-21 konfluen diinfeksi virus ND di dalam 6-well plate, setelah 24 jam infeksi terlihat perubahan morfologi sel atau CPE (Cytopathic Effect) berupa pemendekan sel pada sel BHK-21. Hasil uji HA menujukan perbedaan titer antara hasil inokulasi in-ovo dan in-vitro, masing-masing sebanyak 26 HAU/25 µL dan 22 HAU/25 µL. Deteksi virus ND hasil inokulasi in-vitro menggunakan uji RT-PCR menunjukkan hasil positif dengan munculnya pita DNA pada panjang amplikon 535 bp. Diperlukan pasase bertahap agar titer virus dapat mencapai keadaan maksimal. Virus ND genotipe VII asal Sulawesi Selatan tahun 2019 mampu tumbuh dalam kultur sel BHK-21.

Newcastle Disease virus is a cause of pathogenic disease in poultry, but behind it’s pathogenic nature to poultry, ND virus has an oncolytic effect that can be used as a virotherapy agent. This study aims to examine the growth of the ND virus genotype VII from South Sulawesi in 2019 in BHK-21 cell line. Confluent BHK-21 cells were infected with ND virus in 6-well plate, after 24 hours of infection visible changes in cell morphology or CPE (Cytopathic Effect) in the form of shortening cells in each BHK-21 cells. HA test results showed a difference titer between in-ovo and in-vitro inoculation result, the titer were 26 HAU/25 µL and 22 HAU/25 µL, respectively. Virus detection using the RT-PCR test showed a positive result with the appearance of DNA bands at the amplicon length of 535 bp. Gradual passage is required for the amount of virus titer can reach a maximum state. ND virus genotype VII from South Sulawesi in 2019 was able to growth in BHK-21 cell line.