Moon, Aerie
(Department of Biotechnology, College of Science and Technology, Dongguk University-Gyeongju)
,
Ahn, Jaehyun
(Ourhome Food Research Institute)
,
Choi, Weon Sang
(Department of Biotechnology, College of Science and Technology, Dongguk University-Gyeongju)
오염된 식품으로부터 바이러스를 효과적으로 검출하기 위해서는 식품에 부착된 바이러스를 효과적으로 elute시키는 것이 결정적으로 중요하다. 본 연구에서는 채소류에 공통적으로 적용할 수 있는 elution용액을 찾기 위해 폴리오 바이러스를 인위적으로 오염시킨 1가지 엽채류 (깻잎) 와 3가지 근채류 (당근, 양파, 무) 로부터 바이러스 회수율을 조사한 후, 최적의 바이러스 회수조건을 찾기 위해 기보고된 상추와 양배추의 회수율을 함께 분석하였다. 바이러스의 회수율은 식품의 matrix와 사용된 elution용액의 종류에 따라 차이가 컸으나 0.25M threonine / 0.3M NaCl (pH 9.5) 또는 0.25M glycine / 0.14M NaCl (pH 9.5)을 사용하였을 때 6가지 채소 중 5가지로부터 폴리오바이러스를 효과적으로 elute 할 수 있었다. 0.25M threonine / 0.3M NaCl(pH 9.5)를 노로바이러스 검출에 적용해 본 결과 근채류인 당근보다 엽채류인 깻잎으로부터 노로바이러스 GII를 더 잘 검출할 수 있었다. 이 같은 공통 elution용액을 사용할 경우 다양한 종류의 채소류에 오염된 노로바이러스를 포함한 소화기바이러스의 검출을 용이하게 해 줄 것이다.
오염된 식품으로부터 바이러스를 효과적으로 검출하기 위해서는 식품에 부착된 바이러스를 효과적으로 elute시키는 것이 결정적으로 중요하다. 본 연구에서는 채소류에 공통적으로 적용할 수 있는 elution용액을 찾기 위해 폴리오 바이러스를 인위적으로 오염시킨 1가지 엽채류 (깻잎) 와 3가지 근채류 (당근, 양파, 무) 로부터 바이러스 회수율을 조사한 후, 최적의 바이러스 회수조건을 찾기 위해 기보고된 상추와 양배추의 회수율을 함께 분석하였다. 바이러스의 회수율은 식품의 matrix와 사용된 elution용액의 종류에 따라 차이가 컸으나 0.25M threonine / 0.3M NaCl (pH 9.5) 또는 0.25M glycine / 0.14M NaCl (pH 9.5)을 사용하였을 때 6가지 채소 중 5가지로부터 폴리오바이러스를 효과적으로 elute 할 수 있었다. 0.25M threonine / 0.3M NaCl(pH 9.5)를 노로바이러스 검출에 적용해 본 결과 근채류인 당근보다 엽채류인 깻잎으로부터 노로바이러스 GII를 더 잘 검출할 수 있었다. 이 같은 공통 elution용액을 사용할 경우 다양한 종류의 채소류에 오염된 노로바이러스를 포함한 소화기바이러스의 검출을 용이하게 해 줄 것이다.
The efficient elution of viruses from contaminated food is a critical step for its detection. In this study, conditions optimal for enteric viral recovery from three leafy vegetables (cabbage, lettuce, sesame leaf) and three root vegetables (carrot, onion, mooli) were analyzed to find common buffers...
The efficient elution of viruses from contaminated food is a critical step for its detection. In this study, conditions optimal for enteric viral recovery from three leafy vegetables (cabbage, lettuce, sesame leaf) and three root vegetables (carrot, onion, mooli) were analyzed to find common buffers (statistically not different) applicable to the leafy and root vegetables. Viral recovery varied depending on the food matrices or elution buffers. Buffer solutions containing 0.25M threonine / 0.3M NaCl (pH 9.5) or 0.25M glycine / 0.14M NaCl (pH 9.5) could efficiently recover poliovirus from five out of six vegetables. The threonine buffer was applied to one leafy vegetable (sesame) and one root vegetable (carrot) for genogroup II norovirus (NoV) detection. The detection sensitivity was significantly higher from the leafy vegetable compared to the root vegetable. The use of these common elution buffers should facilitate the detection of low levels of NoV and other enteric viruses in a wide range of vegetables.
The efficient elution of viruses from contaminated food is a critical step for its detection. In this study, conditions optimal for enteric viral recovery from three leafy vegetables (cabbage, lettuce, sesame leaf) and three root vegetables (carrot, onion, mooli) were analyzed to find common buffers (statistically not different) applicable to the leafy and root vegetables. Viral recovery varied depending on the food matrices or elution buffers. Buffer solutions containing 0.25M threonine / 0.3M NaCl (pH 9.5) or 0.25M glycine / 0.14M NaCl (pH 9.5) could efficiently recover poliovirus from five out of six vegetables. The threonine buffer was applied to one leafy vegetable (sesame) and one root vegetable (carrot) for genogroup II norovirus (NoV) detection. The detection sensitivity was significantly higher from the leafy vegetable compared to the root vegetable. The use of these common elution buffers should facilitate the detection of low levels of NoV and other enteric viruses in a wide range of vegetables.
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제안 방법
45 µm porosity; Pall Corporation, USA). All experiments were performed at least three times independently and recovery were calculated based on the titer of the added poliovirus stock (set at 100%) counted on every trial.
Beef extract4,21), amino acid buffer3,8,22), Tris buffer23), amino acid-NaCl buffer3,9,14,19,24), Tris-glycine buffer3,25), or a combination of these buffers6,12,19,26) were examined for their ability to elute virus from produce in several studies. In this study, several buffers including PBS, beef extract, glycine buffers, and threonine buffers were compared for viral recovery from one leafy vegetable (sesame leaf) and three root vegetables (carrot, onion, mooli) (Table 1). The recovery data for the leafy vegetables (cabbage and lettuce) from Moon et al.
2 RT-PCR units, which was determined using serial dilutions of stool samples (data not shown). The level of NoV GII detection for the procedure was determined by inoculating serially-diluted NoV GII on sesame leaves (leafy vegetables) or carrots (root vegetables), processing with 0.25M threonine/0.3M NaCl (pH 9.5), and then examining final RNA concentrates by RT-PCR combined with semi-nested PCR (NoV GII). Poliovirus (10,000 pfu) was used as a positive control to monitor proper processing.
오염된 식품으로부터 바이러스를 효과적으로 검출하기 위해서는 식품에 부착된 바이러스를 효과적으로 elute시키는 것이 결정적으로 중요하다. 본 연구에서는 채소류에 공통적으로 적용할 수 있는 elution용액을 찾기 위해 폴리오 바이러스를 인위적으로 오염시킨 1가지 엽채류 (깻잎) 와 3가지 근채류 (당근, 양파, 무) 로부터 바이러스 회수율을 조사한 후, 최적의 바이러스 회수조건을 찾기 위해 기보고된 상추와 양배추의 회수율을 함께 분석하였다. 바이러스의 회수율은 식품의 matrix와 사용된 elution용액의 종류에 따라 차이가 컸으나 0.
대상 데이터
. The NoV GII used in this study was obtained from the Division of Enteric and Hepatitis Viruses at the National Institute of Health, (Seoul, Korea). The titer of NoV GII stock was 128 RT-PCR units/ml.
성능/효과
5)을 사용하였을 때 6가지 채소 중 5가지로부터 폴리오바이러스를 효과적으로 elute 할 수 있었다. 0.25M threonine / 0.3M NaCl (pH 9.5)를 노로바이러스 검출에 적용해 본 결과 근채류인당근보다 엽채류인 깻잎으로부터 노로바이러스 GII를 더잘 검출할 수 있었다. 이 같은 공통 elution용액을 사용할 경우 다양한 종류의 채소류에 오염된 노로바이러스를 포함한 소화기바이러스의 검출을 용이하게 해 줄 것이다.
본 연구에서는 채소류에 공통적으로 적용할 수 있는 elution용액을 찾기 위해 폴리오 바이러스를 인위적으로 오염시킨 1가지 엽채류 (깻잎) 와 3가지 근채류 (당근, 양파, 무) 로부터 바이러스 회수율을 조사한 후, 최적의 바이러스 회수조건을 찾기 위해 기보고된 상추와 양배추의 회수율을 함께 분석하였다. 바이러스의 회수율은 식품의 matrix와 사용된 elution용액의 종류에 따라 차이가 컸으나 0.25M threonine / 0.3M NaCl (pH 9.5) 또는 0.25M glycine / 0.14M NaCl (pH 9.5)을 사용하였을 때 6가지 채소 중 5가지로부터 폴리오바이러스를 효과적으로 elute 할 수 있었다. 0.
후속연구
5)를 노로바이러스 검출에 적용해 본 결과 근채류인당근보다 엽채류인 깻잎으로부터 노로바이러스 GII를 더잘 검출할 수 있었다. 이 같은 공통 elution용액을 사용할 경우 다양한 종류의 채소류에 오염된 노로바이러스를 포함한 소화기바이러스의 검출을 용이하게 해 줄 것이다.
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