Yüksek İnfektif Titre ve 146S FMD Virus Partikülü Eldesi İçin Uygun BHK-21 Hücre Kültürü Hatlarının Saptanması

Yıl 2021, Cilt: 10 Sayı: 1, 20 - 27, 22.06.2021

Veli Gülyaz , Ahu Kader Kara Neslihan Taşçene , Banu Bayri Özbilğe , Yasemin Gültekin , Mustafa Hasöksüz , Gonca Öztap

https://doi.org/10.31196/huvfd.808775

Öz

Şap Hastalığı (FMD), çift tırnaklı hayvanların en bulaşıcı hastalıklarından biri olup duyarlı hayvanlarda ciddi ekonomik kayba neden olma konusunda büyük bir potansiyele sahiptir. FMD hastalığına karşı mücadelede inaktif FMD aşıları yaygın olarak kullanılmaktadır. Bu çalışma, FMD aşı üretiminde fazla miktarda 146S FMD (O, A ve Asia-1) virüs partikülü içeren virüs süspansiyonu elde etmek için uygun BHK-21 hücre hatlarının tespiti amacıyla yapıldı. Bu amaçla şu anda FMD aşı üretiminde kullanılan BHK-21 An30 hücre kültürüne ek olarak, BHK-21 An73 hücre hatları 20 kez pasajlandı. Her 5 pasajda bir, FMD virüsünün O, A ve Asia-1 serotiplerinin enfektif titresi ve 146S virüs partikül miktarları ölçüldü. Ayrıca, her iki hücre hattının 20. pasaj seviyelerinde karyotip analizi yapıldı ve hücrelerin morfolojik özelliklerinde ve kromozom yapılarında önemli bir değişiklik olmadığı saptandı. FMD virüsünün O, A ve Asia-1 serotiplerinin BHK-21 An73 hücre hattında elde edilen enfektif titre ve 146S virüs partikül miktarlarının, BHK-21 An30 hücre hattında saptanan değerlerden yüksek olduğu belirlendi. FMD virüsünün O, A ve Asia-1 serotiplerinin 146S virüs partikül miktarlarındaki artışların istatistiksel olarak anlamlı olduğu saptandı (p <0.05). Sonuç olarak, FMD virusuna karşı en duyarlı BHK-21 hücre hatlarının önceden belirlenmesi ile aşı üretim prosesinde yüksek titrede FMD 146S virus partikülü elde edilebilecektir. Aşı üretim kapasitesinin artmasından dolayı üreticiye ekonomik olarak önemli katkı sağlayacağı kanaatine varıldı.

Anahtar Kelimeler

BHK-21, Hücre hattı, FMDV, Aşı, Üretim

Destekleyen Kurum

Tarımsal Araştırmalar ve politikalar Genel müdürlüğü (TAGEM)

Proje Numarası

TAGEM/HSGYAD/16/A02/P02/78

Determination of Appropriate BHK-21 Cell Line to Obtain High Infective Titer and 146S FMD Virus Particles

Öz

Foot and mouth disease (FMD) is one the most contagious diseases in cloven-hoofed animals and has a great potential for causing severe economic loss in susceptible animals. In the fight against FMD disease, inactive FMD vaccines are widely used. This study was carried out for the detection of suitable BHK-21 cell lines in order to get virus suspensions containing excess amounts of 146S FMD virus serotypes O, A and Asia-1, for FMD vaccine production. For this purpose, in addition to the BHK-21 An30 cell culture currently used in FMD vaccine production, BHK-21 An73 cell lines were passaged 20 times. In every 5 passages, infective titers and 146S virus particle quantities of FMD virus serotypes O, A and Asia-1 were measured. Additionally, karyotype analysis was performed on both cells at the 20th passage levels and the tests revealed that there was no significant change in the morphological features and chromosomal structures of the cells.
It was determined that the infective titer and 146S virus particle quantities of serotypes O, A and Asia-1 of FMD virus were very high in the BHK-21 An73 cell line than they are available in the BHK-21 An30 cell line. The increases of 146S virus particle quantities of serotypes O, A and Asia-1 of FMD virus were statistically significant (p <0.05).
As a result, high titer FMD 146S virus particles can be obtained in the vaccine production process by pre-determining the BHK-21 cell lines that are most sensitive to the FMD virus. Due to the increase in vaccine production capacity, it was concluded that it would make an important economic contribution to the producer.

Anahtar Kelimeler

BHK-21, FMDV, Vaccine, Production, Cell line

Proje Numarası

TAGEM/HSGYAD/16/A02/P02/78

Kaynakça

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