Sookhee Ha, Gun-Jo Woo, In-Gyun Hwang, Weon Sang Choi
언어
영어(ENG)
URL
https://www.earticle.net/Article/A112614
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원문정보
초록
영어
Low levels of virus contamination and naturally occurring reverse transcription-polymerase chain reaction (RTPCR) inhibitors restrain virus detection in oysters. A rapid and efficient oyster-processing procedure that can be used for sensitive virus detection in oysters was developed. Poliovirus type 1 Sabin strain was used to evaluate the efficacy of virus recovery. The procedure included (a) acid-adsorption and elution with buffers (0.25M glycine-0.14 M NaCl, pH 7.5; 0.25M threonine-0.14M NaCl, pH 7.5); (b) polyethylene glycol (PEG) precipitation; (c) resuspension in Tween 80/Tris solution and chloroform extraction; (d) the second PEG precipitation; (e) viral RNA extraction with TRIzol and isopropanol precipitation; and (f) RT-PCR combined with semi-nested PCR. The overall recovery of elution/concentration was 19.5% with poliovirus. The whole procedure usually takes 19 hr. The overall detection sensitivity was 4 RT-PCR units of genogroup I norovirus (NoV) and 6.4 RT-PCR units of genogroup II Nov/25 g of oysters initially seeded. The virus-detecting method developed in this study should facilitate the detection of low levels of NoV in oysters.
목차
Abstract Introduction Materials and Methods Results and Discussion References
키워드
elutionconcentrationnoroviruspoliovirusoyster
저자
Sookhee Ha [ Department of Biotechnology, Dongguk University ]
Gun-Jo Woo [ Division of Food Bioscience & Technology, Korea University ]
In-Gyun Hwang [ Food Microbiology Division, Korea Food & Drug Administration ]
Weon Sang Choi [ Department of Biotechnology, Dongguk University ]
Corresponding Author