[논문]Bacteriophage MS2를 이용한 소독제 효력시험 확립에 관한 연구

이채홍; 김수희; 한복희; 김영욱; 허문; 정우석

doi:10.7853/kjvs.2021.44.3.149

Bacteriophage MS2를 이용한 소독제 효력시험 확립에 관한 연구
Method development for efficacy testing of veterinary disinfectants using bacteriophage MS2 원문보기

韓國家畜衛生學會誌 = Korean journal of veterinary service, v.44 no.3, 2021년, pp.149 - 155

이채홍 (농림축산검역본부 동물질병관리부 동물약품평가과) , 김수희 (농림축산검역본부 동물질병관리부 동물약품평가과) , 한복희 (농림축산검역본부 동물질병관리부 동물약품평가과) , 김영욱 (농림축산검역본부 동물질병관리부 동물약품평가과) , 허문 (농림축산검역본부 동물질병관리부 동물약품평가과) , 정우석 (농림축산검역본부 동물질병관리부 동물약품평가과)

Abstract ▼ AI-Helper

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 disinfectants 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 biological 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.

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표/그림 (3)

표 Table 1. Characteristics of bacteriophage MS2
그림 Fig. 1. Schematic representation of double agar layer method for disinfectant efficacy test.
그림 Fig. 2. Example of phage lysis plaque phenotypes observed in bacteriophage MS2 onto E. coli C3000 lawn. The presence of phage can be identified as clear zones, or plaques, on a cloudy suspension of bacterial cells growing in the soft agar.

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Bacteriophage MS2를 이용한 소독제 효력시험 확립에 관한 연구
Method development for efficacy testing of veterinary disinfectants using bacteriophage MS2 원문보기

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Bacteriophage MS2를 이용한 소독제 효력시험 확립에 관한 연구 Method development for efficacy testing of veterinary disinfectants using bacteriophage MS2 원문보기

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Bacteriophage MS2를 이용한 소독제 효력시험 확립에 관한 연구
Method development for efficacy testing of veterinary disinfectants using bacteriophage MS2 원문보기