关键词: 新型鸭呼肠孤病毒, SYBR GreenⅠ, 实时荧光定量RT\, PCR

Abstract:  A pair of specific primers targeted to gene S1 of the novel duck reovirus （NDRV） was designed in this study， and a fluorescent quantitative RT\|PCR （qRT\|PCR） assay based on SYBR Green Ⅰ fluorescent was also developed for novel duck reovirus （NDRV） detection. The standard curve was plotted based on the linear relationship between the amount of plasmid DNA and the cycle threshold.The sensitivity test showed that the detection limit of qRT\|PCR was about 15 copies for the cDNA of target gene， and the minimum detectable amount of the virus was 05 TCID50. This method only detected NDRV， but not the duck tembusu virus （DTMUV）， A\|type duck hepatitis virus （DHV\|1）， C\|type duck hepatitis virus （DHV\|3）， muscovy duck reovirus （MDRV）， egg drop syndrome virus （EDSV）， duck Newcastle disease virus （NDV）， duck plague virus （DPV）， and Riemerella anatipestifer （RA）. The method had good repeatability with intra\|assay variation coefficients of 032%-091% and inter\|assay variation coefficients of 103%-151%. The feces of artificially infected ducks were detected by the method， it was found that 2-12 d after attack was the detoxification period， of which 3-6 d was the peak. It took only 4 hours from receiving the clinical samples to obtaining the detection results by this method， which was more sensitive than conventional RT\|PCR assay.

Key words:  novel duck reovirus, SYBR GreenⅠ, qRT\, PCR