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RT-PCR 및 Nested PCR기법을 이용한 동물용 생 바이러스 백신 중 Bovine Viral Diarrhea Virus (Pestivirus) 오염 신속검출법 확립
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  • RT-PCR 및 Nested PCR기법을 이용한 동물용 생 바이러스 백신 중 Bovine Viral Diarrhea Virus (Pestivirus) 오염 신속검출법 확립
저자명
안동준,전우진,정병열,이철현,박봉균,정갑수,김병한,An. Dong-Jun,Jeon. Woo-Jin,Jung. Byeong-Yeal,Yi. Chul-Hyun,Park. Bong-Kyun,Chung. Gab-Soo,Kim. Byoun
간행물명
Journal of bacteriology and virology : JBV
권/호정보
2005년|35권 3호|pp.273-281 (9 pages)
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대한미생물학회
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이 논문은 한국과학기술정보연구원과 논문 연계를 통해 무료로 제공되는 원문입니다.
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기타언어초록

Bovine viral diarrhea virus(BVDV) of the genus Pestivirus is hewn as a common contaminant of cell culture-derived vaccines. Hog cholera virus(HCV), which is also of the genus Pestivirus and an important livestock disease in Korea, is recognized as a potential contaminant of vaccines produced in porcine cells. However, it is difficult for the National Biological Assays of korea to adequately detect contamination of these agents in biological products. for these reasons, we established rapid and sensitive methods fir the detection of BVDV and HCV contamination in cell cultures and veterinary biologicals by using RT-PCR and nested PCR assays. We designed a Pestivirus primer amplifying 152 bp to detect both BVDV and HCV and a common primer amplifying 237 bp to detect only BVDV. Also, for the differentiation between BVDV type 1 and type 2, nested PCR was conducted using the amplified 237bp PCR product, to amplify 179bp in BVDV type 2 genome. The sensitivity of the PCR using common primer for the detection of BVDV was $400;TCID_{50}/ml$. All 6 strains of Korean BVDV isolates, 5 vaccines strains and the standard strain NADL could be detected. No reactions were observed ohen testing 5 types of viruses infecting pigs (HCV, TGEV, PEDV, JEV, PRRSV), 4 types infecting cattle (Akabane virus, BEFV, BCV, BRV) and 4 types infecting cats (FIP, FPL, FCV, FVR). Using this RT-PCR assay, commercial vaccines were tested and, 55 lots from 12 vaccine companies, were negative for BVDV contaminations. Same results were obtained by the immunoflourescence assay. The newly developed PCR or RT-PCR assays can be used as rapid, reliable, sensitive, and simple methods for the detection of BVDV (Pestivirus) in cell cultures, master seeds, and live viral vaccines.