Establishment and Preliminary Application of a Multiple PCR Assay for NDRV, NGPV, and DTMUV Detections

  Objective  A PCR method for simultaneous detection of new-type duck reovirus (NDRV), novel goose parvovirus (NGPV), and duck tembusu virus (DTMUV) was developed and tested for clinic applications.

  Method  Separate sets of specific primers were designed based on the conserved regions of NDRV, NGPV, and DTMUV genomes. The target specific amplifications of NDRV, NGPV, and DTMUV were in the regions of 594 bp, 467 bp, and 328 bp, respectively. The specificity, sensitivity, and repeatability of the assay method were determined prior to a trial determination on clinic specimens.

  Result  The newly developed multiplex PCR methodology delivered highly repeatable results and showed a high specificity on the 3 viruses with negative results on other commonly found duck pathogens. The detection sensitivity on NDRV was 8.80×10⁴ copies·μL⁻¹, 4.03×10⁴ copies·μL⁻¹ on NGPV, and 2.15×10⁴ copies·μL⁻¹ on DTMUV. On 167 clinical samples a perfect 100% total coincidence rate between the new and conventional PCR methods were achieved in detecting the 3 viruses.

  Conclusion   The newly established multiplex PCR assay was specific, sensitive, and repeatable in simultaneously detecting NDRV, NGPV, and DTMUV. It was considered adequate for clinic applications.