Bacteriophage MS2를 이용한 소독제 효력시험 확립에 관한 연구
Method development for efficacy testing of veterinary disinfectants using bacteriophage MS2
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In virucidal efficacy testing, the chemical inactivation cannot be determined for all viruses due to the difficulties or the inability to culture sufficiently or the risk of exposure to the viruses. Therefore, disinfectants against these viruses could be evaluated by different methods and surrogate viruses are used as alternative. In this study we developed a method for efficacy testing of veterinary disin-fectants using one of the candidate surrogate viruses, bacteriophage MS2, as part of the research on the selection of surrogate viruses for efficiency of efficacy testing of veterinary disinfectants. This method is based on the Animal and Plant Quarantine Agency (APQA) guidelines for efficacy testing of veterinary disinfectants. Bacteriophage and disinfectant are reacted in suspension in accordance with the APQA guidelines and then a newly established double agar layer method is applied for the efficacy test. The double agar layer method is summarized as follows: 1) The bottom agar with 1.5% agar is boiled and cooled before poured into petri dishes at volume of 20 mL, and dried under bio-logical safety cabinet. 2) The top agar with 0.7% agar is boiled and kept at 50℃ before E. coli culture was seeded. 3) The serially diluted bacteriophage MS2-disinfectant mixtures 0.05 mL and E. coli host 0.01 mL (OD600 0.2~0.3) are mixed with 5 mL of top agar and incubate them at 50℃ for 5 min for reaction. 4) The resulting mixture is poured over top of a bottom agar plate and rocked sufficiently to ensure that the top agar covers the entire surface of the bottom agar. 5) The double agar layer is then placed under biological safety cabinet to allow the agar layer to solidify and subsequently incubated at 37℃ for 24 hr. 6) Following incubation, the plates may be inspected for plaques and record results.
Abstract
In virucidal efficacy testing, the chemical inactivation cannot be determined for all viruses due to the difficulties or the inability to culture sufficiently or the risk of exposure to the viruses. Therefore, disinfectants against these viruses could be evaluated by different methods and surrogate viruses are used as alternative. In this study we developed a method for efficacy testing of veterinary disin-fectants using one of the candidate surrogate viruses, bacteriophage MS2, as part of the research on the selection of surrogate viruses for efficiency of efficacy testing of veterinary disinfectants. This method is based on the Animal and Plant Quarantine Agency (APQA) guidelines for efficacy testing of veterinary disinfectants. Bacteriophage and disinfectant are reacted in suspension in accordance with the APQA guidelines and then a newly established double agar layer method is applied for the efficacy test. The double agar layer method is summarized as follows: 1) The bottom agar with 1.5% agar is boiled and cooled before poured into petri dishes at volume of 20 mL, and dried under bio-logical safety cabinet. 2) The top agar with 0.7% agar is boiled and kept at 50℃ before E. coli culture was seeded. 3) The serially diluted bacteriophage MS2-disinfectant mixtures 0.05 mL and E. coli host 0.01 mL (OD600 0.2~0.3) are mixed with 5 mL of top agar and incubate them at 50℃ for 5 min for reaction. 4) The resulting mixture is poured over top of a bottom agar plate and rocked sufficiently to ensure that the top agar covers the entire surface of the bottom agar. 5) The double agar layer is then placed under biological safety cabinet to allow the agar layer to solidify and subsequently incubated at 37℃ for 24 hr. 6) Following incubation, the plates may be inspected for plaques and record results.
참고문헌 (21)
- 농림축산검역본부, "소독제 효력시험지침", 농림축산검역본부 고시 제2018-16호, ~ (2018)
- 배경선, "하수 처리 과정의 염소 소독에 대한 여러 박테리오파지들의 저항성 평가; 물 재이용 과정의 안전성 관리를 위한 바이러스 지표미생물의 개발", 상하수도학회지, 30(3), pp.285~291 (2016), 대한상하수도학회
- 서영석, "전해 염소수", 자외선 결합 시스템을 이용한 병원성 미생물의 불활성화 키네틱스 평가, 2019, pp.33(5), 379~388[DOI]
- Adcock NJ, "The use of bacteriophages of the family Cystoviridae as surrogates for H5N1 highly pathogenic avian influenza viruses in persistence and inactivation studies", J Environ Sci Health A Tox Hazard Subst Environ Eng, 44, pp.1362~1366 (2009)
- Aquino de Carvalho NA, "Evaluation of Phi6 persistence and suitability as an enveloped virus surrogate", Environ Sci Technol, 51, pp.8692~8700 (2017)
- Buckley D, "Efficacy of novel aqueous photo-chlorine dioxide against a human norovirus surrogate, bacteriophage MS2 and Clostridium difficile endospores, in suspension, on stainless steel and under greenhouse conditions", J Appl Microbiol, 130, pp.1531~1545 (2020)
- Cho M, "Different inactivation behaviors of MS-2 phage and Escherichia coli in TiO2 photocatalytic disinfection", Appl Environ Microbiol, 71, pp.270~275 (2005)
- Cormier J, "A double layer plaque assay using spread plate technique for enumeration of bacteriophage MS2", J Virol Methods, 196, pp.86~92 (2014)
- Eggers M, "The European tiered approach for virucidal efficacy testing – rationale for rapidly selecting disinfectants against emerging and re-emerging viral diseases", Euro Surveill, 26 (2021)[DOI]
- Gallandat K, "Selection of a Biosafety Level 1 (BSL-1) surrogate to evaluate surface disinfection efficacy in Ebola outbreaks: Comparison of four bacteriophages", PLoS ONE, 12, pp.e0177943~ (2017)[DOI]
- Kampf G, "Efficacy of three ethanol-based hand rubs against feline calicivirus, a surrogate virus for norovirus", J Hosp Infect, 60, pp.144~149 (2005)
- Lillehaug, D, "An improved plaque assay for poor plaque-producing temperate lactococcal bacteriophages", J Appl Microbiol, 83, pp.85~90 (1997)
- Morin T, "Comparison of the virucidal efficacy of peracetic acid, potassium monopersulphate and sodium hypochlorite on bacteriophages P001 and MS2", J Appl Microbiol, 119, pp.655~665 (2015)
- Prodělalová J, "Virucides in apiculture : persistence of surrogate enterovirus under simulated field conditions", Pest Manag Sci, 73, pp.2544~2549 (2017)
- Sanders ER, "Aseptic Laboratory Techniques: Plating Methods", J Vis Exp, 63, pp.e3064~ (2012)[DOI]
- Steinmann J, "Surrogate viruses for testing virucidal efficacy of chemical disinfectants", J Hosp Infect, 56, pp.S49~S54 (2004)
- Turgeon N, "Comparison of five bacteriophages as models for viral aerosol studies", Appl Environ Microbiol, 80, pp.4242~4250 (2014)
- "Method 1601: Male-specific (F+) and somatic coliphage in water by two-step enrichment procedure" (2001)[URL]
- Wyrzykowska-Ceradini B, "The use of bacteriophage MS2 for the development and application of a virucide decontamination test method for porous and heavily soiled surfaces", J Appl Microbiol, 127, pp.1315~1326 (2019)
- Ye Y, "Survivability, partitioning, and recovery of enveloped viruses in untreated municipal wastewater", Environ Sci Technol, 50, pp.5077~5085 (2016)