Determination of Appropriate BHK-21 Cell Line to Obtain High Infective Titer and 146S FMD Virus Particles
Yıl: 2021 Cilt: 10 Sayı: 1 Sayfa Aralığı: 20 - 27 Metin Dili: İngilizce DOI: 10.31196/huvfd.808775 İndeks Tarihi: 29-07-2022
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 greatpotential for causing severe economic loss in susceptible animals. In the fight against FMD disease, inactive FMD vaccinesare widely used. This study was carried out for the detection of suitable BHK-21 cell lines in order to get virus suspensionscontaining excess amounts of 146S FMD virus serotypes O, A and Asia-1, for FMD vaccine production. For this purpose, inaddition to the BHK-21 An30 cell culture currently used in FMD vaccine production, BHK-21 An73 cell lines were passaged20 times. In every 5 passages, infective titers and 146S virus particle quantities of FMD virus serotypes O, A and Asia-1 weremeasured. Additionally, karyotype analysis was performed on both cells at the 20th passage levels and the tests revealedthat 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 werevery high in the BHK-21 An73 cell line than they are available in the BHK-21 An30 cell line. The increases of 146S virusparticle 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 theBHK-21 cell lines that are most sensitive to the FMD virus. Due to the increase in vaccine production capacity, it wasconcluded that it would make an important economic contribution to the producer.
Anahtar Kelime: Yüksek İnfektif Titre ve 146S FMD Virus Partikülü Eldesi İçin Uygun BHK-21 Hücre Kültürü Hatlarının Saptanması
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 Kelime: Belge Türü: Makale Makale Türü: Araştırma Makalesi Erişim Türü: Erişime Açık
- Abbas F, Khan FA, Ahmad F, Hussain A, Ahmad M et al, 2011: Production of foot and mouth disease virus vaccine (O Type) on BHK-21. cell line, Iğdır Univ J Ins Sci Tech, 1(2), 155-159.
- Aktas S, Samuel AR, 2000: Identification of antigenic epitopes on the foot and mouth disease virus isolate O1/Manisa/Turkey/1969 using monoclonal antibodies. Rev Sci Tech, 19, 744-753.
- Alexandersen S, Zhang Z, Donaldson AI, Garland AJM. 2003: The pathogenesis and diagnosis of foot and mouth disease. J Comp Path, 129, 1-36.
- Ali SM, Ismail AH, Soliman EM, Hanaa AM, 2013: Studies on growth kinetics of the FMDV serotype SAT-2 Egyptian strain in cell culture. J Vet Adv, 3(2), 92-97.
- Archettı I, Amadorı M, Donn A, Salt J, Lodettı E, 1995: Detection of foot-and-mouth disease virus-infected cattle by assessment of antibody response in oropharyngeal fluids. J Clin Microbiol, 33(1), 79-84.
- Baer A, Kehn-Hall K, 2014: Viral concentration determination through plaque assays. J Vis Exp, 93, 1-10.
- Bartelling SJ, 2002: Development and performance of inactivated vaccines against foot and mouth disease. Rev Sci Tech Off Int Epiz, 21, 577-588.
- Bartelling SJ, Vreeswijk J, 1991: Developments in footand-mouth disease vaccines. Vaccine, 9(2), 5-88.
- Berg K, Mohr J, 1963: Genetics of the Lp system. Acta Genet, 13(4), 349-360.
- Callens M, De Clercq K, 1997: Differentiation of the seven serotypes of foot-and-mouth disease virus by reverse transcriptase polymerase chain reaction. J Virol Methods, 67, 35-44.
- Chapman WG, Stewart DL, 1962: Growth of a cloned strain of hamster kidney cells in suspended cultures and their susceptibility to the virus of foot-andmouth disease. Nature. 195, 1163.
- Czelleng F, Zsitvay K, Egyhazi ZS, Baranyi M, Fazekas A, 1987: Comparative analysis of BHK-21 cell lines of virüs strains of foot-and-mounth disease. Arch Exper Vet Med, 41(6), 701-796.
- Çokçalışkan C, Türkoğlu T, Sareyyüpoğlu B, Uzunlu E, Babak A., et al, 2016: Qs-21 enhances the early antibody response to oil adjuvant foot and mouth disease vaccine in cattle. Clin Exp Vac Res, 5,138- 147.
- Fransis BMJ, Fry CM, Rowlands DJ, Brown F, Bitte JL et al, 1987: Immunological priming with synthetic peptides of foot-and-mouth disease. Immunology, 61(1), 1-6.
- Freshney RI, 2010: A manual of basic technique and specialized application. Wiley-Blackwell, A John Wiley&Sons, Inc. Pub.
- Grubman MJ, Baxt B, 2004: Foot and mouth disease. Clin Microbiol Rev, 17(2), 465-493.
- Gunetti M, Castiglia S, Rustichelli D, Mareschi K, Sanavio F et al, 2012: Validation of analytical methods in GMP: the disposable fast read 102W device, an alternative practical approach for cell counting. J Trans Med, 31,10-112.
- Harmsen MM, Fitjen HPD, Westra DF, Coco-Martin JM, 2011: Effect of thiomersal on dissociation of intact (146s) foot-and-mouth disease virions into 12s particles as assessed by novel elisas specific for either 146s or 12s particles. Vaccine, 29, 2682-2690.
- Jerome P, Howard LB, 1964: Production and purification of milligram amounts of foot-and-mouth disease virus from baby hamster kidney cell cultures. J Appl Microbiol, 12 (4), 368-373.
- Nawaz Z, Siddique AB, Zahoor MA, Bilal Aslam, B, Zahoor MK et al, 2019: Detectıon of foot and mouth dısease vırus sheddıng in mılk of apparently healthy buffaloes and cattle of Punjab, Pakistan. Buffalo Bull, 38(2), 255-261.
- OIE, 2017: OIE Manual of Diagnostic Tests and Vaccines for Terrestrial Animals. Terrestrial Manual: foot and mouth disease: Chapter 3.1.8, (May), 433-464.
- Özbilge BB., Gülyaz V, Taşçene N, Yılmaz Ş, Gültekin Y et al, 2020. Determination of the effects of enrofloxacin, linco-spectinand florfenicol antibiotics on BHK-21 cell culture and FMD 146s virus particlesınfective titers. Etlik Vet Mikrobiyol Derg, 31(1), 7- 19.
- Paton DJ, Taylor G, 2011: Developing vaccines against foot-and-mouth disease and some other exotic viral diseases of livestock. Phil Trans R Soc B, 366(1579), 2774–2781.
- Rahman SU, Rabbani M, Sahidullah K, Muhammed Z, 2007: Studies on in vitro culture characteristics of adherent baby hamster kidney-21(BHK-21) cell line. Int J Agri Biol, 9(6), 821-826.
- Rweyemamu MM, Umehara O, Giorci W, Medeiros R, Lucca DN et al, 1989: Effect of formaldehyde and binary ethyleneimine (BEI) on the integrity of foot and mouth disease virus capsid. Rev Sci Tech Off Int Epiz, 8(3), 747-764.
- Sareyyüpoğlu B, Gülyaz V, Cokçalışkan C, Ünal Y, Çökülgen T, et al, 2019: Effect of fmd vaccination schedule of dams on the level and duration of maternally derived antibodies. Vet Immunol Immunopathol, Vol, 217.
- Shirai J, Chatchawanchonteera A, Sinsuwongwat W, Makarasen P, Sugimura T, 1990: Estimation of 146s particles in foot and mouth disease virus (FMDV) vaccine by using computer analysing system. Jpn J Vet Sci, 52(3), 621-630.
- Xiao Y, Chen HY, Wang Y, Yin B, Lv C et al, 2016: Largescale production of foot-and-mouth disease virus (serotype Asia-1) VLP vaccine in escherichia coli and protection potency evaluation in cattle. BMC Biotechnology, 16: 56.
- Van Wezel AL, 1967: Growth of cell-strains and primary cells on microcarriers in homogeneous culture. Nature, 216; 64.
| APA | GÜLYAZ V, KARA A, Taşçene N, bayri ozbilge b, GULTEKİN Y, HASOKSÜZ M, ÖZTAP ÖZEN G (2021). Determination of Appropriate BHK-21 Cell Line to Obtain High Infective Titer and 146S FMD Virus Particles. , 20 - 27. 10.31196/huvfd.808775 |
| Chicago | GÜLYAZ Veli,KARA Ahu Kader,Taşçene Neslihan,bayri ozbilge banu,GULTEKİN YASEMİN,HASOKSÜZ MUSTAFA,ÖZTAP ÖZEN Gonca Determination of Appropriate BHK-21 Cell Line to Obtain High Infective Titer and 146S FMD Virus Particles. (2021): 20 - 27. 10.31196/huvfd.808775 |
| MLA | GÜLYAZ Veli,KARA Ahu Kader,Taşçene Neslihan,bayri ozbilge banu,GULTEKİN YASEMİN,HASOKSÜZ MUSTAFA,ÖZTAP ÖZEN Gonca Determination of Appropriate BHK-21 Cell Line to Obtain High Infective Titer and 146S FMD Virus Particles. , 2021, ss.20 - 27. 10.31196/huvfd.808775 |
| AMA | GÜLYAZ V,KARA A,Taşçene N,bayri ozbilge b,GULTEKİN Y,HASOKSÜZ M,ÖZTAP ÖZEN G Determination of Appropriate BHK-21 Cell Line to Obtain High Infective Titer and 146S FMD Virus Particles. . 2021; 20 - 27. 10.31196/huvfd.808775 |
| Vancouver | GÜLYAZ V,KARA A,Taşçene N,bayri ozbilge b,GULTEKİN Y,HASOKSÜZ M,ÖZTAP ÖZEN G Determination of Appropriate BHK-21 Cell Line to Obtain High Infective Titer and 146S FMD Virus Particles. . 2021; 20 - 27. 10.31196/huvfd.808775 |
| IEEE | GÜLYAZ V,KARA A,Taşçene N,bayri ozbilge b,GULTEKİN Y,HASOKSÜZ M,ÖZTAP ÖZEN G "Determination of Appropriate BHK-21 Cell Line to Obtain High Infective Titer and 146S FMD Virus Particles." , ss.20 - 27, 2021. 10.31196/huvfd.808775 |
| ISNAD | GÜLYAZ, Veli vd. "Determination of Appropriate BHK-21 Cell Line to Obtain High Infective Titer and 146S FMD Virus Particles". (2021), 20-27. https://doi.org/10.31196/huvfd.808775 |
| APA | GÜLYAZ V, KARA A, Taşçene N, bayri ozbilge b, GULTEKİN Y, HASOKSÜZ M, ÖZTAP ÖZEN G (2021). Determination of Appropriate BHK-21 Cell Line to Obtain High Infective Titer and 146S FMD Virus Particles. Harran Üniversitesi Veteriner Fakültesi Dergisi, 10(1), 20 - 27. 10.31196/huvfd.808775 |
| Chicago | GÜLYAZ Veli,KARA Ahu Kader,Taşçene Neslihan,bayri ozbilge banu,GULTEKİN YASEMİN,HASOKSÜZ MUSTAFA,ÖZTAP ÖZEN Gonca Determination of Appropriate BHK-21 Cell Line to Obtain High Infective Titer and 146S FMD Virus Particles. Harran Üniversitesi Veteriner Fakültesi Dergisi 10, no.1 (2021): 20 - 27. 10.31196/huvfd.808775 |
| MLA | GÜLYAZ Veli,KARA Ahu Kader,Taşçene Neslihan,bayri ozbilge banu,GULTEKİN YASEMİN,HASOKSÜZ MUSTAFA,ÖZTAP ÖZEN Gonca Determination of Appropriate BHK-21 Cell Line to Obtain High Infective Titer and 146S FMD Virus Particles. Harran Üniversitesi Veteriner Fakültesi Dergisi, vol.10, no.1, 2021, ss.20 - 27. 10.31196/huvfd.808775 |
| AMA | GÜLYAZ V,KARA A,Taşçene N,bayri ozbilge b,GULTEKİN Y,HASOKSÜZ M,ÖZTAP ÖZEN G Determination of Appropriate BHK-21 Cell Line to Obtain High Infective Titer and 146S FMD Virus Particles. Harran Üniversitesi Veteriner Fakültesi Dergisi. 2021; 10(1): 20 - 27. 10.31196/huvfd.808775 |
| Vancouver | GÜLYAZ V,KARA A,Taşçene N,bayri ozbilge b,GULTEKİN Y,HASOKSÜZ M,ÖZTAP ÖZEN G Determination of Appropriate BHK-21 Cell Line to Obtain High Infective Titer and 146S FMD Virus Particles. Harran Üniversitesi Veteriner Fakültesi Dergisi. 2021; 10(1): 20 - 27. 10.31196/huvfd.808775 |
| IEEE | GÜLYAZ V,KARA A,Taşçene N,bayri ozbilge b,GULTEKİN Y,HASOKSÜZ M,ÖZTAP ÖZEN G "Determination of Appropriate BHK-21 Cell Line to Obtain High Infective Titer and 146S FMD Virus Particles." Harran Üniversitesi Veteriner Fakültesi Dergisi, 10, ss.20 - 27, 2021. 10.31196/huvfd.808775 |
| ISNAD | GÜLYAZ, Veli vd. "Determination of Appropriate BHK-21 Cell Line to Obtain High Infective Titer and 146S FMD Virus Particles". Harran Üniversitesi Veteriner Fakültesi Dergisi 10/1 (2021), 20-27. https://doi.org/10.31196/huvfd.808775 |
