Differential diagnosis of CSFV wild strain and vaccine strain by the Fluorescent Quantitative RT-PCR Technique. Issue 2 (March 2019)

• Record Type: Journal Article
• Authors: Huang, Jing; Li, Kun; Zhang, Zhengyi; Li, Chong; Liu, Huan; Wang, Qinfu
• Is Part Of: Journal of physics. Volume 1176:Issue 2(2019)
• Publication Date: 2019-03
• Subjects: Physics -- Congresses; 530.5
• DOI: 10.1088/1742-6596/1176/2/022022
• Languages: English
• ISSNs: 1742-6588

Abstract

Abstract: The purpose of this study was to establish a rapid method for distinguishing the virulent strains of CSFV from vaccine strains. In GeneBank, sequence alignment was performed using DNAstar's Meglign. A specific amplification primer was designed according to the independent T rich insertion sequence in 3'-UTR of CSFV vaccine strain. The fluorescent quantitative RT-PCR technique showed a high specificity and sensitivity. The CSFV Shimen strain, CSFV live vaccine cell line origin, CSFV Thiverval strain and CSFV live vaccine spleen-lymph tissue-origin showed specific band, while the other BVDV C24V strain, PRRSV vaccine Tianjin strain, JEV vaccine 14-14-2 strain, and PEDV-TGEV-PoRV live vaccine strains showed no specific bands. The length of PCR product for CSFV wild strain and vaccine strain are 99 bp and 115-140 bp, respectively. The 10 5 TCID50 of the vaccine was diluted into 10 0, 10 −1, 10 −2, 10 −3, 10 −4, 10 −5 and 10 −6 . The maximum times of dilution could be detected by the real-time RT-PCR was 10 −6 . So, the sensitivity reached up to 0.1 TCID50. The fluorescent quantitative RT-PCR technique was set up successfully for differential diagnosis of CSFV wild strain and vaccine strain, which is prerequisite for eradication of CSF in swine farm.