Development and Application of the Nested PCR Assay for Duck Flavivirus

作者：ZHANG Lin; HU Bei xia; LU Mao yang; YANG Shao hua; XU Chuan tian; ZHANG Xiu mei

来源：Animal Husbandry and Feed Science, 2013, (01): 3-7.

• Flavivirus
• Universal primers
• Nested PCR
• Duck
• Sensitivity

[Objective] Aim to establish a kind of efficient detection method for duck flavivirus(DFV). [Method] The method of nested PCR based on flavivirus universal primers which were designed according to the GeneBank flavivirus gene sequence. [Result] The degenerate universal primers Flav P1-Flav P4 were designed by genome comparison to target NS5 gene conserved area. The system could only amplify flavivirus purpose gene and the sensitivity was 90 copies/μL, higher than the ordinary PCR 1 000 times. Homology and evolutionary analysis showed that duck flavivirus belonged to mosquito-born flavivirus, NTAV group, similar with Tembusu and BYD virus. [Conclusion]Primers of the nested PCR system had good universality and specificity and method had high sensitivity. This system successfully detected flavivirus and clarified the evolution station of DFV.

• 出版日期2013-2-15
• 单位中国农业大学