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논문 기본 정보

자료유형
학위논문
저자정보

이희민 (전남대학교, 전남대학교 대학원)

지도교수
김두운
발행연도
2014
저작권
전남대학교 논문은 저작권에 의해 보호받습니다.

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이 논문의 연구 히스토리 (3)

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Current molecular methods that include PCR have been used to detect norovirus and hepatitis A virus in many food samples. However, the protocols the require removal of PCR inhibitors and time-consuming concentration steps to separate virus from analyte for rapid and sensitive detection of norovirus and hepatitis A virus. We developed combination of immunomagnetic separation (IMS) and a quantum dots (QDs) assay to detect norovirus and hepatitis A virus eluted from fresh lettuce with Tris buffer containing 1% beef extract (pH 9.5). IMS facilitated viral precipitation with a 10-min incubation, whereas virus concentration using polyethylene glycol (PEG) requires more than 3 h and an additional high-speed centrifugation step to precipitate virus before reverse transcription PCR (RT-PCR) analysis. The detection limits of IMS-QDs for NoV and HAV were 1RT-PCR unit/ml and 10 TCID50/mL, which was as similar result using RT-PCR combined with PEG or IMS, respectively. The results suggest that a fluorescence assay based on IMS and QDs is valid for detecting NoV and HAV qualitatively. The application of IMS-QDs assay completed viral detection within a 1 hour, but not possible within this time using PEG combined with RT-PCR. In conclusion, the IMS-QDs assay is both rapid and efficient to detect NoV and HAV at low concentration in agricultural products.

목차

Abstract 3
Abbreviations 5
Ⅰ. Introduction 6
Ⅱ. Materials and Methods 9
1. Virus 9
2. Virus extraction and concentration 9
3. RT-PCR 10
4. Detection of NoV or HAV with a fluorescent quantum dot probe 11
5. Statistical Analysis 13
Ⅲ. Results 14
1. Optimization of specific NoV detection method using IMS 14
2. Comparison of two concentration methods for detecting NoV 17
3. NoV detection limits using IMS and QDs 17
4. Optimization of the specific HAV detection method using IMS 21
5. Comparison of two concentration methods for detecting HAV 21
6. HAV detection limits using IMS and QDs 24
Ⅳ. Discussion 26
Ⅴ. References 28
Ⅵ. Abstract (in Korean) 33

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