Prophylactic Control Of Mycoplasma Contamination In Starting Biological Materials Used In Viral Vaccine Production | ||||
| Journal of Applied Veterinary Sciences | ||||
| Article 5, Volume 4, Issue 1, April 2019, Page 30-34 PDF (361.01 K) | ||||
| Document Type: Original Article | ||||
| DOI: 10.21608/javs.2019.62673 | ||||
 View on SCiNiTO | ||||
| Author | ||||
Ahmed F. Soudy 
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| Veterinary Serum and Vaccine Research Institute, Abassia, Cairo, Egypt | ||||
| Abstract | ||||
| Mycoplasma contamination remains a major concern in the biopharmaceutical industry especially in tissue culture based viral vaccine and its presence and/or its endotoxin-like metabolites in the final products can result in pyrogenic responses ranging from fever and chills, to irreversible and fatal septic shock. This study was conducted by in vitro screening of mycoplasma in the different ingredients used in production of Foot and mouth Disease (FMD) vaccine using Polymerase Chain Reaction (PCR) using universal primers that are specific to the 16S rRNA region. Tested items include growth media, cell lines, trypsin, seed virus and working virus. Also the study evaluates the inhibitory effect of different concentrations of neomycin, kanamycin, gentamycin, polymyxin B and ciprofloxacin on mycoplasma contaminated cell lines. Our results showed that the prepared growth media, trypsin, seed virus as well as working virus were mycoplasma free and three tested cell lines were also free while another two lines were mycoplasma positive. The mycoplasma positive cell line are poorly grown in comparison with the free line using the same growth media and the virus yield from the apparently normal contaminated line was very low. Ciprofloxacin can be used for treating valuable cell line after 12 days in 25mg/L and after 18 days in 10mg/L. Ciprofloxacin plus regular antibiotic may keep the line sterile for prolonged time but treatment of contaminated cell line is not advisable. So, prophylactic control by strict personal hygiene and personal protective equipment (PPE) and adopt appropriate aseptic techniques is the core solution. | ||||
| Keywords | ||||
| FMD; Mycoplasma; PCR. Vaccine; VSVRI | ||||
| References | ||||
|
ANTHONY O. OLARERIN-GEORGE JOHN B. AND HOGENESCH 2015. Assessing the prevalence of mycoplasma contamination in cell culture via a survey of NCBI's RNA-seq archive Nucleic Acids Research, Volume 43, Issue 5, 11 March 2015, Pages2535–542,https://doi.org/10.1093/nar/gkv136.
BARILE MF, and ROTTEM S .1993. Mycoplasmas in cell cultures. In: Kahane I, Adoni A, editors. Rapid diagnosis of mycoplasmas. New York: Plenum Press. pp. 155-193.
CORD C. UPHOFF, SABINE-A. DENKMANN, AND HANS G. DREXLER 2012. Treatment of Mycoplasma Contamination in Cell Cultures with Plasmocin. Hindawi Publishing Corporation Journal of Biomedicine and Biotechnology Volume 2012, Article D 267678,8pages doi:10.1155/2012/267678.
DAVIS, J.M., 2002. Basic Cell Culture, 2nd Ed, Pp: 167–70. Oxford University, Press U.K DREXLER HG, and UPHOFF CC .2002. Mycoplasma contamination of cell cultures: incidence, sources, effects, detection, elimination, prevention. Cytotechnology.39:75. doi: 10.1023/A: 1022913015916.
DSMZ.2012. German Collection of Microorganisms and Cell Cultures: Catalogue of Human and Animal Cell Lines, 2012.
GENZEL Y, FISCHER M, and REICHL U. 2006. Serum-free influenza virus production avoiding washing steps and medium exchange in large-scale microcarrier culture. Vaccine. 24:3261–72.
GERLIC M, HOROWITZ J, FARKASH S, AND HOROWITZ S. 2007. The inhibitory effect of Mycoplasma fermentans on tumor necrosis factor (TNF)-alpha-induced apoptosis resides in the membrane lipoproteins. Cell Microbiol. 9:142-153.
GRUBMAN MJ, BAXT B.2004. Foot-and-mouth disease. Clin Microbiol Rev. 2004;17: 465–93.
HASSAN AI.2016. Effect of different culture systems on the production of foot and mouth disease trivalent vaccine. Vet World. 2016;9:32–7.
MANISCHEWITZ JE, YOUNG BG, and BARILE MF. 1975. The effect of mycoplasmas on replication and plaquing ability of Herpes simplex virus. Proc. Soc. Exp. Biol. Med. 148: 859-863.
NIKFARJAM. L, AND FARZANEH. P, 2012. Prevention and detection of mycoplasma contamination in cell culture. Cell J. 13:4: 203-12.
PARIDA S. 2009. Vaccination against foot-and-mouth disease virus: strategies and effectiveness. Expert Rev. Vaccines, 8:347–65.
TIMENETSKY J, SANTOS L, and BUZINHANI M. 2006. Detection of multiple mycoplasma infection in cell cultures by PCR. Braz J Med Biol Res, 39:907–914. doi: 10.1590/S0100-879X. YOUNG, L. SUNG J, STACEY G, and. MASTERS J. R.2010. Detection of Mycoplasma in cell cultures, Nature protocols, vol. 5, no. 5, pp. 929–934, 2010.
UPHOFF CC, DREXLER HG. 2002. Comparative PCR analysis for detection of mycoplasma infections in continuous cell lines. In Vitro Cell Dev Bio Anim. 2002;38:79–85. doi: 10.1290/1071-2690. VAHID MOLLA KAZEMIHA, MOHAMMAD ALI SHOKRGOZAR,MOHAMMAD REZA ARABESTANI,MORTEZA SHOJAEI MOGHADAM, SHAHRAM AZARI,SUSAN MALEKI, AMIR AMANZADEH, MAHMOOD JEDDI TEHRANI, and FAZEL SHOKRI, 2009. PCR-based detection and eradication of mycoplasmal infections from various mammalian cell lines: a local experience. Dec; 61(3): 117–124. VOLOKHOV DV, GRAHAM LJ, BRORSON KA, and CHIZHIKOV VE. 2011. Mycoplasma testing of cell substrates and biologics: Review of alternative non- microbiological techniques. Molecular and cellular probes 25: 69-77, 2011.
YUEYANG LAI, XUEBO XU, RUIYING YAN, and ZICHUN H.U.A .2019. Evaluation of mycoplasma removal reagents using qPCR-based quantification. Analytical Biochemistry. Volumes 564–565, 1 January 2019, Pages 88-95. | ||||
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