Abstract
In order to compare protocols for inactivation of viruses potentially present in biological specimens, three different model viruses were treated in bovine serum by two different inactivation methods: samples were subjected either to chemical inactivation with ethylenimine (El) at concentrations of 5 and 10 mM at 37 degrees C for periods up to 72 h or to electron-beam irradiation in frozen and liquid form with doses varying between 11 and 46 kGy. The chemical inactivation resulted in nonlinear tailing curves in a semilogarithmic plot of virus titer versus inactivation time showing non-first-order kinetics with respect to virus titer. The time for inactivation of 7 log10 units of porcine parvovirus (PPV) was about 24 h for both El concentrations, whereas 5 log10 units of bovine viral diarrhea virus (BVDV) was inactivated in 2 h for both El concentrations and 6 log10 units of porcine enterovirus (PEV) was inactivated within 3 h. The inactivation with electron-beam irradiation resulted in almost linear curves in a semilogarithmic plot of virus titer versus irradiation dose, reflecting a first-order inactivation. The rate of inactivation was almost twice as fast in the liquid samples compared to the rate in frozen ones, giving values of the doses needed to reduce virus infectivity 1 log10 unit for inactivation of PPV of 11.8 and 7.7 kGy for frozen and liquid samples, respectively, whereas the corresponding values for BVDV were 4.9 and 2.5 kGy, respectively, and those for PEV were 6.4 and 4.4 kGy, respectively. The nonlinear inactivation with El makes it impossible to extrapolate the curves beyond the virus detection limit and thereby predict the necessary time for complete inactivation, i.e., to a level beyond the detection limit, of virus in a given sample. The first-order inactivation obtained with electron-beam irradiation makes such a prediction possible and justifiable. The two methods are discussed with respect to their different kinetics and applicability under different circumstances and criteria for inactivation, and considerations for choice of method are discussed.
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Selected References
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- Bahnemann H. G. Binary ethylenimine as an inactivant for foot-and-mouth disease virus and its application for vaccine production. Arch Virol. 1975;47(1):47–56. doi: 10.1007/BF01315592. [DOI] [PubMed] [Google Scholar]
- Bahnemann H. G. The inactivation of foot-and-mouth disease virus by ethylenimine and propylenimine. Zentralbl Veterinarmed B. 1972 Jul;20(5):356–360. doi: 10.1111/j.1439-0450.1973.tb01136.x. [DOI] [PubMed] [Google Scholar]
- Bauer K. Die Inaktivierung des Maul- und Klauenseuche (MKS)-Virus durch Aethylaethylenimin und die Eignung des inaktivierten Virus zur Impfstoffherstellung. Zentralbl Bakteriol Orig. 1970 Apr;213(3):285–297. [PubMed] [Google Scholar]
- Blackburn N. K., Besselaar T. G. A study of the effect of chemical inactivants on the epitopes of Rift Valley fever virus glycoproteins using monoclonal antibodies. J Virol Methods. 1991 Aug;33(3):367–374. doi: 10.1016/0166-0934(91)90036-y. [DOI] [PubMed] [Google Scholar]
- Cunliffe H. R. Inactivation of foot-and-mouth disease virus with ethylenimine. Appl Microbiol. 1973 Nov;26(5):747–750. doi: 10.1128/am.26.5.747-750.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Elliott L. H., McCormick J. B., Johnson K. M. Inactivation of Lassa, Marburg, and Ebola viruses by gamma irradiation. J Clin Microbiol. 1982 Oct;16(4):704–708. doi: 10.1128/jcm.16.4.704-708.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
- HUBER W. Cold sterilization by electron beam as a possible tool for the inactivation of the virus of homologous serum jaundice in plasma. Ann N Y Acad Sci. 1952 Sep;55(3):536–542. doi: 10.1111/j.1749-6632.1952.tb26574.x. [DOI] [PubMed] [Google Scholar]
- Hassanain M. M. Preliminary findings for an inactivated African horsesickness vaccine using binary ethyleneimine. Rev Elev Med Vet Pays Trop. 1992;45(3-4):231–234. [PubMed] [Google Scholar]
- House C., House J. A., Yedloutschnig R. J. Inactivation of viral agents in bovine serum by gamma irradiation. Can J Microbiol. 1990 Oct;36(10):737–740. doi: 10.1139/m90-126. [DOI] [PubMed] [Google Scholar]
- Kyvsgaard N. C., Lind P., Preuss T., Kamstrup S., Lei J. C., Bøgh H. O., Nansen P. Activity of antibodies against Salmonella dublin, Toxoplasma gondii, or Actinobacillus pleuropneumoniae in sera after treatment with electron beam irradiation or binary ethylenimine. Clin Diagn Lab Immunol. 1996 Nov;3(6):628–634. doi: 10.1128/cdli.3.6.628-634.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Latarjet R. Interaction of radiation energy with nucleic acids. Curr Top Radiat Res Q. 1972 Jul;8(1):1–38. [PubMed] [Google Scholar]
- Mahnel H., Stettmund von Brodorotti H., Ottis K. Empfindlichkeit von Viren gegen Gammastrahlen. Zentralbl Bakteriol B. 1980 Feb;170(1-2):57–70. [PubMed] [Google Scholar]
- May E. M., Hunt D. C., Sloggem J. A normal-phase high-performance liquid chromatographic assay for aziridine residue in trientine dihydrochloride. J Pharm Biomed Anal. 1987;5(1):65–70. doi: 10.1016/0731-7085(87)80009-2. [DOI] [PubMed] [Google Scholar]
- Sun I. L., Gustafson D. P., Scherba G. Comparison of pseudorabies virus inactivated by bromo-ethylene-imine, 60Co irradiation, and acridine dye in immune assay systems. J Clin Microbiol. 1978 Nov;8(5):604–611. doi: 10.1128/jcm.8.5.604-611.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Thomas F. C., Davies A. G., Dulac G. C., Willis N. G., Papp-Vid G., Girard A. Gamma ray inactivation of some animal viruses. Can J Comp Med. 1981 Oct;45(4):397–399. [PMC free article] [PubMed] [Google Scholar]