关键词: 水禽细小病毒, 检测方法, SYBR Green I, 荧光定量PCR

Abstract: In order to establish a rapid detection method for waterfowl parvoviruses （WPV）, specific primers were designed within the conserved SF3 region of the NS gene of waterfowl parvoviruses based on sequence alignment results, and a SYBR Green I fluorescence quantitative PCR universal detection method was established. The amplification efficiency （E） of this method was 90.0%, the correlation coefficient （R²） was 0.99, and the standard curve equation was y=-3.607x+38.77;except for WPV with an S-shaped amplification curve, the newcastle disease virus （NDV）, H9 subtype avian influenza virus （H9 AIV）, duck tembusu virus （DTMUV）, duck hepatitis A virus （DHAV）, duck enteritis virus （DEV）, and duck reovirus （DRV） samples did not show an S-shaped positive amplification curve;the coefficient of variation （CV） within a batch was 0.15% to 0.23%, and the coefficient of variation between batches was 0.09% to 0.28%. The results indicated that the SYBR Green I fluorescence quantitative PCR detection method had good repeatability, high sensitivity, and strong specificity. The clinical sample testing results showed that the coincidence rate between SYBR Green I fluorescence quantitative PCR and conventional PCR was 98.4%, and the sensitivity was 1 000 times higher than that of conventional PCR. The SYBR Green I fluorescence quantitative PCR detection method could not only qualitatively detect WPV, but also quantitatively detect it. It could be used for WPV purification detection in duck and goose breeding farms, as well as for rapid detection of WPV in large clinical samples.

Key words: waterfowl parvoviruses, detection method, SYBR Green I, fluorescence quantitative PCR