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논문 기본 정보
- 자료유형
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- 한국미생물생명공학회 Journal of Microbiology and Biotechnology Journal of Microbiology and Biotechnology 제27권 제3호
- 발행연도
- 2017.1
- 수록면
- 471 - 479 (9page)
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초록· 키워드
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White spot syndrome virus (WSSV) is a major threat to the shrimp farming industry and so far there is no effective therapy for it, and thus early diagnostic of WSSV is of great importance. However, at the early stage of infection, the extremely low-abundance of WSSV DNA challenges the detection sensitivity and accuracy of PCR. To effectively detect low-abundance WSSV, here we developed a pre-amplification PCR (pre-amp PCR) method to amplify trace amounts of WSSV DNA from massive background genomic DNA. Combining with normal specific PCR, 10 copies of target WSSV genes were detected from ~1010 magnitude of backgrounds. In particular, multiple target genes were able to be balanced amplified with similar efficiency due to the usage of the universal primer. The efficiency of the pre-amp PCR was validated by nested-PCR and quantitative PCR, and pre-amp PCR showed higher efficiency than nested-PCR when multiple targets were detected. The developed method is particularly suitable for the super early diagnosis of WSSV, and has potential to be applied in other low-abundance sample detection cases.
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참고문헌 (23)
참고문헌 신청
Andersson D / 2015 / Properties of targeted preamplification in DNA and cDNA quantification / Expert Rev. Mol. Diagn 15 : 1085 ~ 1100
Arandjelovic M / 2009 / Two-step multiplex polymerase chain reaction improves the speed and accuracy of genotyping using DNA from noninvasive and museum samples / Mol. Ecol.
Chou H / 1995 / Pathogenicity of a baculovirus infection causing white spot syndrome in cultured penaeid shrimp in Taiwan / Dis. Aquat. Organ 23 : 165 ~ 173
Escobedo-Bonilla CM / 2008 / A review on the morphology, molecular characterization, morphogenesis and pathogenesis of white spot syndrome virus / J. Fish Dis 31 : 1 ~ 18
Hameed AS / 2003 / Experimental infection of twenty species of Indian marine crabs with white spot syndrome virus (WSSV) / Dis. Aquat. Organ 57 : 157 ~ 161
Arandjelovic M / 2009 / Two-step multiplex polymerase chain reaction improves the speed and accuracy of genotyping using DNA from noninvasive and museum samples / Mol. Ecol.
Chou H / 1995 / Pathogenicity of a baculovirus infection causing white spot syndrome in cultured penaeid shrimp in Taiwan / Dis. Aquat. Organ 23 : 165 ~ 173
Escobedo-Bonilla CM / 2008 / A review on the morphology, molecular characterization, morphogenesis and pathogenesis of white spot syndrome virus / J. Fish Dis 31 : 1 ~ 18
Hameed AS / 2003 / Experimental infection of twenty species of Indian marine crabs with white spot syndrome virus (WSSV) / Dis. Aquat. Organ 57 : 157 ~ 161
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