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A Pair of Novel Primers for universal detection of the NS1 Gene from Various Bluetongue Virus Serotypes

Virologica Sinica 2008年第1期23卷 68-72页

作者： Hui-qiong YIN Gai-ping ZHANG Hong ZHANG Jin-gang ZHANG Viral Safety Laboratory of National Center of Biomedical Analysis Institute of Transfusion Medicine The Academy of Military Medical Sciences Beijing 100850 China Henan Key Laboratory of Animal Immunology Henan 450002 China

Twenty five serotypes of Bluetongue virus (BTV) have been identified worldwide. Rapid and reliable methods of virus universal detection are essential for fighting against bluetongue (BT). We have therefore developed and evaluated a pair of primers which can detect various serotypes of BTV by RT-PCR. Analysis of the viral protein 7 (VP7) and the non-structural protein (NS1) gene from different serotypes of BTV by DNAstar showed that the 5' end of the NS1 gene is the most conserved region. The primer pairs (P1 and P2) were designed based on the highly conserved region of NS1. The novel primers were evaluated by detecting BTV serotypes 1, 3, 5, 8, 10, 11, 21 and 22. The specificity of the primers was estimated by comparing to gene sequences of viruses published in GenBank, and further assessed by detecting BTV serotype 1-12 and Epizootic hemorrhagic disease virus (EHDV) serotype 1-4. The sensitivity and repeatability of PCR with the novel primers were evaluated by successfully detecting the recombinant plasmid pGEM-T121 containing the diagnosed nucleotide sequence. Our results suggest that these unique primers can be used in high throughout and universal detection of the NS1 gene from various BTV serotypes.

关键词： RT-PCR Bluetongue virus (BTV) NS1 universal detection

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