A Rapid And Quantitative Method To Detect Pseudorabies Virus

PRV is an important virus which acutely threaten the breeding.The virus particle has high resistance to circumstances.gE gene is the pecant gene of PRV and being the deleted gene of gene deleted vaccines.The method contains two pairs of primers of gE,gB gene each and an internal dual-labeled fluorogenic probe of gE gene.We use Flurogenic quantitative PCR combines PE7700 sequence detection system to first established a rapid and reproducible method for assessment of PRV loads in animals and products.The sensitivity of the assay was 101copies per reaction.The assay is linear within 7-log dynamic range. The sensitivity was obviously higher than gE-SYBR green PCR and conventional PCR. We compared the charisteristics and applicability of Taqman PCR and SYBR green PCR.gE-MGB-Taqman PCR was used to detected 18 frigorific tissue samples,the positive rate is 15/16 (other 2deserted) which had the 100% coincidence with Serum neutralization and Cell MTT chromatometry.The closed-tube format of the assay minimized the risk of contamination of subsequent reaction and the assay can be performed in 2h or less.