Abstract:
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.
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