Académique Documents
Professionnel Documents
Culture Documents
S. Specter D. Jeffries
Virucidal agents may inactivate viruses either because of their physical or chemical properties. These agents generally are effective on viruses outside of their host cells. The ability of these agents to inactivate virus is very closely associated with the conditions under which they are used. Depending on the agent, this may include temperature, pH, moisture content of the sample, concentration and composition of the virus, concentration of the agent, and time of exposure. Thus, viruses kept at room temperature will become inactivated more rapidly than those held at 4~ Likewise some viruses maintained at neutral pH may be inactivated less rapidly than at either lower or higher pH (Lancz 1976), other viruses such as enteroviruses may be stable at highly acidic pH. Some viruses are also known to lose infectivity as the sample in which they are collected dries (Parker et al 1944). Thus, virus maintained in mucus containing secretions on an environmental surface is likely to retain infectivity longer than the same amount of virus in saliva, since the latter will dry more quickly (Dunham 1977). An important factor in the effectiveness of disinfection is the virus titer present at the outset of addition of the disinfectant. The loss of infectivity over time is such that if a sample that is treated with a virucidal agent is held for 15 min it may lose 50% of its infectivity, after 30 min 90% is lost, and by 60 min there is a 99% loss. If a sample contains 1000 infectious particles per milliliter at outset, after 15 min 0.1 ml will contain approximately 5 infectious units, whereas after 60 min 0.1 ml Virology Methods Manual ISBN 0-12-465330-8 has a 10% chance of containing infectious material. By contrast, if the original concentration of virus is 106 particles per milliliter, then even a 99% inactivation after 1 h may leave sufficient infectious material to cause infection; i.e. 1000 infectious units will be present in 0.1 ml. The ability of various agents to inactivate viruses is also dependent on the composition of the virus. Many viruses contain a lipid envelope that is sensitive to lipid solvents such as ether, chloroform or detergents, whereas naked (non-enveloped) viruses are resistant to such agents (Dunham 1977). Finally, the nature of the disinfection agents can determine the extent of inactivation. This will depend upon whether the agent is physical or chemical and if it is organic or inorganic. It should be noted that other organic material in solutions containing viruses may inhibit the virucidal effects of many disinfectants. Wiedbrauk and Johnston (1993) have delineated which compounds are particularly useful for viruses that are commonly encountered both in the laboratory and on environmental surfaces. Inactivation of commonly used disinfectants is shown in Table 18.1. The methods for testing virucidal agents are well delineated (Koski and Chen 1977).
Protein
Hard Water
Detergents
++++ + + + +
, . . , . . m
+ + + + +
.
C C -..
* Alcohols are extremely poor in penetrating proteinaceous material and are, therefore, of very limited use as disinfectants. C, Cationic detergents chlorine and bromine (Brown et a11963; Clarke and Kabler 1964; Lund 1964). The most readily available antiviral for use is common household bleach (sodium hypochlorite), which usually can be used as a 10% solution (the solution must contain 10,000 ppm available chlorine), to inactivate many enveloped and naked viruses. The bleach is capable of destroying nucleic acids, making it a universally effective disinfectant for viruses. Thus, it is the recommended disinfectant for routine use in the clinical laboratory, the physician's office, and any setting where concerns about viral contamination exist. Its ability to inactivate nucleic acids makes household bleach an ideal agent for decontamination of a laboratory where polymerase chain reactions are performed, and carry over of nucleic acids is a concern (Prince and Andrews 1992). Chlorine and bromine are useful for the inactivation of viruses in swimming pools when used at about 0.5 parts per million. Iodine also can be used as a disinfectant of contaminated water supplies (Dunham and MacNeal 1944; Kabler 1962). Ozone is considered to be safer and more effective than chlorine for the disinfection of water, but is not practical for widespread use due to cost (Sliter 1974). Similarly, hydrogen peroxide can be used to inactivate viruses. However, the presence of other organic materials in fluids will compete with the viruses and block the effects of the H202. Acid solutions, such as 1 N HCI, and alkaline compounds, such as 2% NaOH are also useful disinfectants for inactivation of viruses. Metals and metal salts also have antiviral properties. Copper salts, mercury, potassium 354 permanganate and silver nitrate have been used as virucidal agents (Dunham 1977).
Physical agents
Elevated temperature has a distinct antiviral effect, increasing the rate of virus inactivation. The inactivation rate due to high temperature varies for different viruses and can be changed greatly by the pH of a solution. The presence of a protein stabilizer such as serum decreases the ability of heat to inactivate virus in a suspension. Most viruses lose some infectivity when heated at 56~ for 30 min. All viruses are destroyed by appropriate autoclaving, 121~ under 15 psi pressure for 15 min. Ultraviolet radiation is also a highly effective virucidal physical agent, as long as it is of suitable wavelength and intensity. Wavelengths of approximately 250 nm have been demonstrated to be effective against influenza virus (Dunham 1977). The effectiveness is significantly diminished when the virus suspension is moved away from the source of radiation. In addition, ultraviolet rays are more effective through air than water, and particulate matter (dust particles or salt crystals) can further decrease effectiveness.
Summary
There are a wide variety of methods that can be used for sterilization, disinfection and antisepsis. When possible, the recommended method for sterilization is steam heating using an autoclave. An alternative effective method of limiting contamination is to use disposable sterile items (needles, syringes, pipettes, tubes, flasks, etc.) that come in a sealed sterile container. Such items, if not autoclavable, can be sterilized using y-irradiation or ethylene oxide gas after packaging, to achieve sterilization. The recommended disinfectants for decontamination of surfaces or solutions containing viruses are bleach or gluteraldehyde. Instruments that are reused should be cleaned with detergents that will remove organic matter, prior to disinfection with these substances, as such material promotes survival of viruses.
Disinfection
355
References
Brown JR, McLean DM, Nixon MC (1963) Can J Pub Hlth 54: 267-270. Clarke NA, Kabler PW (1954) Am J Hyg 59: 119127. Cox HR, van der Scheer J, Aiston S, Bohnel E (1947) J Immunol 56: 149-166. Dunham WB (1977)In: Disinfection, Sterilization and Preservation, 2nd Edn, Block SS (Ed.) Lea and Febiger, Philadelphia, pp. 426-441 Dunham WB, MacNeal WJ (1944) J Immunol 49: 123-128. Hanson PJV, Gor D, Jeffries DJ, Collins JV (1989) Brit Med J 298: 862-864. Kabler PW (1962) Ann Rev Microbiol 16: 127-140. Klein, M Deforest A (1963) Soap Chem Spec 39: 7072. Koski TA, Chen JHS (1977) In: Disinfection, Sterilization and Preservation 2nd Edn, Block SS (Ed.) Lea and Febiger, Philadelphia, pp. 116-134. Lancz GJ (1976) Virology 75: 488-491. Lund E (1964) Am J Hyg 80: 1-10. McCrea JF (1960) Ann NY Acad Sci 83: 692-705. National Formulary, 12th Ed. (NF XlI) (1965) American Pharmaceutical Assoc, Washington, pp. 3637. Parker ER, Dunham WB, MacNeal WJ (1944) J Lab Clin Med 29: 37-42. Prince AM, Andrus L (1992) BioTechniques 12: 358360. Sliter JT (1974) J Water Pollut Control Fed 46: 4-6. Wiedbrauk DL, Johnston SLG (1993) Manual of Clinical Virology, Raven Press, New York, 273 pp.
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Chapter 18