Skripsi

Pengaruh Konsentrasi Virus Dan TPCK-trypsin terhadap Propagasi Virus Influenza H1N1 pada Sel Madin-Darby Canine Kidney (MDCK) = Effect of Virus and TPCK-Trypsin Concentration on the Propagation of Influenza H1N1 Virus in Madin-Darby Canine Kidney (MDCK) Cells.

Latar Belakang Influenza H1N1 adalah virus yang dapat menyebabkan infeksi saluran napas musiman maupun pandemi. Propagasi virus penting untuk diagnosis dan produksi vaksin. Metode propagasi menggunakan telur ayam memiliki keterbatasan waktu dan risiko mutasi sehingga kultur sel MDCK menjadi alternatif. Namun, propagasi di kultur sel MDCK masih belum bisa menghasilkan titer yang optimal. Oleh karena itu, penelitian ini dilakukan untuk mencari kondisi propagasi yang lebih baik dengan menyesuaikan konsentrasi virus awal dalam HA unit dan konsentrasi TPCK-trypsin. Metode Penelitian eksperimental dilakukan dengan variasi pengenceran konsentrasi virus H1N1 1/10 (8 HAU), 1/100 (0,8 HAU), 1/1000 (0,08 HAU) serta konsentrasi TPCK-trypsin 1,5 dan 2 µg/ml. Virus H1N1 dipropagasi di sel MDCK selama 48 jam di dalam inkubator dengan suhu 37oC dan kadar CO2 5%. Kemudian titernya diukur menggunakan uji hemaglutinasi. Analisis statistik dilakukan untuk mengevaluasi pengaruh konsentrasi virus dan TPCK-trypsin terhadap propagasi virus. Hasil Konsentrasi awal virus memengaruhi titer HA. Konsentrasi virus tinggi (1/10) menunjukkan penurunan titer pasca propagasi akibat ko-infeksi partikel non-infeksius, tetapi lebih tinggi dibanding konsentrasi rendah (1/100 dan 1/1000) karena virus belum mencapai titer maksimum saat panen. Perubahan konsentrasi TPCK-trypsin tidak signifikan memengaruhi titer HA. Kesimpulan Konsentrasi awal virus berperan penting dalam propagasi H1N1 di sel MDCK, sedangkan penurunan konsentrasi TPCK-trypsin tidak mengurangi replikasi virus secara signifikan. Hasil ini dapat dijadikan dasar optimalisasi propagasi H1N1 di sel MDCK untuk tujuan diagnosis dan produksi vaksin.
Kata Kunci: hemaglutinasi, influenza H1N1, konsentrasi, MDCK, trypsin.


Introduction Influenza H1N1 can cause seasonal and pandemic respiratory infections. Virus propagation is essential for diagnosis and vaccine production. While egg-based propagation has time limitations and mutations risk, MDCK cell culture is a viable alternative. However, propagation in MDCK cell culture has not yet been able to produce optimal titers. Therefore, this study was conducted to find better propagation conditions by adjusting the initial virus concentration in HA units and the TPCK-trypsin concentration. Method This experimental study tested various dilutions of H1N1 virus at concentrations of 1/10 (8 HAU), 1/100 (0.8 HAU), 1/1000 (0.08 HAU) and TPCK-trypsin concentrations of 1,5 and 2 µg/ml. The H1N1 virus was propagated in MDCK cells for 48 hours in an incubator at at 37oC and 5% CO2. The virus titer then was measured by hemagglutination assay. Statistical analysis evaluated the effects of virus and TPCK-trypsin on virus propagation. Results Initial virus concentration affected the HA titer. High virus concentration (1/10) showed a post-propagation decrease due to co-infection with non-infectious particles, but remained higher than lower concentrations (1/100 and 1/1000) as the virus didn’t reached maximum titer at harvest. TPCK-trypsin concentrations change didn’t significantly affect HA titer. Conclusion Initial virus concentration plays a crucial role in H1N1 propagation in MDCK cells, while reducing TPCK-trypsin does not significantly reduce viral replication. These findings can guide the optimization of H1N1 propagation in MDCK cells for diagnostic and vaccine production purposes.
Keywords: concentration, hemagglutination, influenza H1N1, MDCK, trypsin