DETERMINATION OF GROWTH CHARACTERISTICS OF THE FOOT AND MOUTH VIRUSES (A, O, ASIA-1) IN BHK-21 AN30 AND BHK-21 AN73 CELL CULTURES

Year 2017, Volume: 2 Issue: 1, 1 - 5, 30.04.2017

Neslihan Taşçene , Banu Bayri Özbilge Sadık Onur Karaçam Veli Gülyaz Mustafa Hasöksüz

Abstract

Bu çalışmada, BHK-21 An₇₃ hücre kültürü ile
BHK-21 An₃₀ hücre kültürlerinin üreme hızı, miktarı ve virus titresi
üzerine etkilerinin araştırılması amaçlandı. Bu amaçla BHK-21 An₃₀ ve
BHK-21 An₇₃ süspanse hücre kültürlerinde, hücrelerin üreme
kinetikleri belirlendi. Bu hücre kültürlerinde A TUR 11, O TUR 07 ve Asia-1/11
şap aşı suşlarının üretimi gerçekleştirildi. BHK-21 An₃₀ hücre
kültürlerinin 7 günlük inkubasyon periyodu esnasında monolayer formda 4. günde
1,3x10⁶/ml hücre sayısıyla, süspanse formda ise 4. günde 4,1x10⁵
hücre sayısı ile pik noktaya ulaşırken BHK-21 An₇₃  kültürlerinde ise monolayer formda 6. günde
2,2x10⁶/ml hücre sayısına ulaşılırken, süspanse formda 3. günde 2,8
x10⁵ hücre sayısına ulaştığı belirlendi.  Süspanse BHK-21 An₃₀ hücre
kültürlerinde A, O ve Asia-1 aşı virusunun üretimi sonrası 146S değerleri
sırasıyla ortalama; 0,51 μg/ml, 0,18 μg/ml ve 0,16 μg/ml olarak saptanırken
enfektif titreleri ortalama 6,87 pfu/ml, 6,22 pfu/ml ve 6,49 pfu/ml olarak
belirlendi.

Süspanse BHK-21 An₇₃ hücre kültürlerinde A,
O ve Asia-1 aşı virusunun üretimi sonrası değerleri sırasıyla ortalama; 2,11
μg/ml, 2,59 μg/ml ve 0,53 μg/ml olarak saptanırken enfektif titreleri ortalama
6,99 pfu/ml, 7,82 pfu/ml ve 6,37 pfu/ml olarak tespit edildi.

Sonuç olarak, aşı üretiminde kullanılan A, O ve Asia-1
aşı viruslarının BHK-21 An₇₃ hücre kültüründe daha yüksek 146S ve
enfektif titrede üreme gösterdiği ve bundan dolayı şap aşısı üretiminde BHK-21
An₇₃ hücre kültürlerinin kullanılmasının gerek üretim zamanı gerekse
maliyet olarak BHK-21 An₃₀ hücre kültürlerinden daha uygun olduğu
kanaatine varılmıştır. 

Keywords

Şap Hastalığı, Şap Virusu suşları, BHK-21 hücre kültürü

DETERMINATION OF GROWTH CHARACTERISTICS OF THE FOOT AND MOUTH VIRUSES (A, O, ASIA-1) IN BHK-21 AN30 AND BHK-21 AN73 CELL CULTURES

Abstract

In
this study, it was aimed to investigate BHK-21An₇₃ and BHK-21An₃₀
growth rate of cell cultures and the effects on foot and mouth disease (FMD)
vaccine virus strains titers. For this purpose, growth rate of suspended cell
cultures of BHK-21An₇₃ and BHK-21An₃₀ were determined. A
TUR 11, O TUR 07 and Asia-1/11 strains of foot and mouth disease (FMD) vaccine
virus strains were produced separately on BHK-21An₃₀ and BHK-21An_73.
BHK-21An₃₀ cell amounts during seven incubation days of
reached to peak level in monolayer form at the 4th day with 1,3x10⁶/ml
cell numbers, in suspension form at the 4th day with 4,1x10⁵ cell
numbers. BHK-21An₇₃ reached to peak level in monolayer form at the
6th day 2,2x10⁶/ml cell numbers while in suspension form at the 3rd
day with 2,8 x10⁵ cell numbers. After the production of A, O and
Asia-1 vaccine viruses, in suspension BHK-21An₃₀ cell cultures
average of 146S values respectively; 0,51 μg/ml, 0,18 μg/ml and 0,16 μg/ml were
detected while infective titers were determined as average 6,87 (Plaque Forming
Units) pfu/ml, 6,22 pfu/ml and 6,49 pfu/ml. After the production of A, O and
Asia-1 vaccine viruses, in suspended BHK-21An₇₃ cell cultures
average of 146S values respectively; 2,11 μg/ml, 2,59 μg/ml and 0,53 μg/ml were
detected while infective titers were determined as average 6,99 pfu/ml, 7,82
pfu/ml and 6,37 pfu/ml. As a result, in the production of A, O and Asia-1
vaccine viruses in BHK-21An_73, resulted in higher 146S values and
higher infective titers than BHK-21An_30. It is concluded that for
FMD vaccine production the using of BHK-21An₇₃ cell culture is more
appropriate for both of cost of manufacture and productive time.

Keywords

Foot and Mouth Disease, FMDV vaccine strains, BHK-21 cell culture

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