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Volume 27 Issue 2
Mar.  2012
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ZHANG Lin, HU Bei-xia, YAN Shi-gan, YANG Shao-hua, XU Chuan-tian, ZHANG Xiu-mei. Development and Application of the Nested PCR Assay for Duck Flavivirus[J]. Fujian Journal of Agricultural Sciences, 2012, 27(2): 124-129.
Citation: ZHANG Lin, HU Bei-xia, YAN Shi-gan, YANG Shao-hua, XU Chuan-tian, ZHANG Xiu-mei. Development and Application of the Nested PCR Assay for Duck Flavivirus[J]. Fujian Journal of Agricultural Sciences, 2012, 27(2): 124-129.
shu

Development and Application of the Nested PCR Assay for Duck Flavivirus

  • 1.

    Institute of Animal Science and Veterinary Medicine, Shandong Academy of Agricultural Science, Jinan, Shandong 250100, China;

  • 2.

    Shandong Key Lab of Animal Disease Control and Breeding, Jinan, Shandong 250100, China

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  • Received Date: April 01, 2011
  • Revised Date: November 18, 2011
    Graphical Abstract
    • Abstract
  • Flavivirus universal primer pairs and nested PCR were developed as an efficient method to detect flavivirus, especially new type flavivirus. Four primers Flav P1-Flav P4 located in conserved region of the NS5 gene were designed after alignment of the flavivirus genome and then nested PCR assay was developed for duck flavivirus. Nested PCR was 1 000 times more sensitive than the conventional PCR and it was only able to amplify the flavivirus with the sensibility of 90 copiesL-1. Phylogenetic analysis showed that duck flavivirus belongs to mosquito-born flavivirus, NTAV group, similar with Tembusu and BYD virus. The clinical samples detection showed that the universal primers and nested PCR were highly specific and sensitive. In addition, the station of evolution of duck flavivirus was clarified.
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    • References (12)
  • 马秀丽,于可响,高凤,等.一株鸭黄病毒的生物血特性研究// 山东畜牧兽医学会禽病学专业委员会第二届禽病学术研讨会, 2011:221-224.
    唐熠,刁有祥,高绪慧,等. 鸭坦布苏病毒逆转录半套式PCR检测方法的建立和应用//山东畜牧兽医学会禽病学专业委员会第二届禽病学术研讨会,2011:209-212.
    SHERYL L MS, NAOMI L F, PAUL J W, et al. Universal primers that amplify RNA from all three flavivirus subgroups[J].Virology Journal, 2008, 5:16.
    KNOTH K, ROBERDS S, POTEET C, et al. Highly degenerate, inosine-containing primers specifically amplify rare cDNA using the polymerase chain reaction[J].Nucleic Acids Research, 1988, 16:10932.
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