Key result
Multiplex PCR simultaneously detects seven duck-infecting viruses with a limit of 10,000 DNA copies/μL.
Why the study?
Major viruses causing harm and economic losses in ducks require a rapid, specific, and sensitive method for simultaneous detection.
The established m-PCR method is feasible for the rapid and specific simultaneous detection of seven duck-infecting viruses.
Answers are grounded in this paper and cite their sources.
Supports multiplex detection of seven duck viruses; leaves open field validation before diagnostic adoption.
Major viruses, including duck-origin avian influenza virus, duck-origin Newcastle disease virus, novel duck parvovirus, duck hepatitis A virus, duck Tembusu virus, fowl adenovirus, and duck enteritis virus, pose great harm to ducks and cause enormous economic losses to duck industry. This study aims to establish a multiplex polymerase chain reaction (m-PCR) method for simultaneous detection of these seven viruses. Specific primers were designed and synthesized according to the conserved region of seven viral gene sequences. Then, seven recombinant plasmids, as the positive controls, were reconstructed in this study. Within the study, D-optimal design was adopted to optimize PCR parameters. The optimum parameters for m-PCR were annealing temperature at 57 °C, Mg 2+ concentration at 4 mM, Taq DNA polymerase concentration at 0.05 U/μL, and dNTP concentration at 0.32 mM. With these optimal parameters, the m-PCR method produced neither cross-reactions among these seven viruses nor nonspecific reactions with other common waterfowl pathogens. The detection limit of m-PCR for each virus was 1 × 10 4 viral DNA copies/μL. In addition, the m-PCR method could detect a combination of several random viruses in co-infection analysis. Finally, the m-PCR method was successfully applied to clinical samples, and the detection results were consistent with uniplex PCR. Given its rapidity, specificity, sensitivity, and convenience, the established m-PCR method is feasible for simultaneous detection of seven duck-infecting viruses and can be applied to clinical diagnosis of viral infection in ducks.
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Yao et al. (2019) studied Viral infection in ducks. Multiplex polymerase chain reaction (m-PCR) method vs. Uniplex PCR was evaluated on Simultaneous detection of seven duck-infecting viruses. A multiplex PCR method was successfully developed for the simultaneous detection of seven duck-infecting viruses with a detection limit of 1 × 10^4 viral DNA copies/μL.
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