Key result
Droplet digital PCR detects PCV-2 with ~4-fold greater sensitivity than TaqMan real-time PCR.
Why the study?
Sensitive detection of Porcine circovirus-2 is important for surveillance of postweaning multisystemic wasting syndrome, and a more precise detection method is needed.
Does droplet digital PCR improve the sensitivity of detecting Porcine circovirus-2 compared to TaqMan real-time PCR?
Does droplet digital PCR improve the sensitivity of detecting Porcine circovirus-2 compared to TaqMan real-time PCR?
Effect estimate: 4-fold greater sensitivity
Droplet digital PCR provides a highly sensitive and specific platform for detecting Porcine circovirus-2 viral loads, outperforming standard TaqMan real-time PCR.
May aid PCV-2 surveillance in swine; leaves open need for field validation versus standard assays.
Sensitive detection of Porcine circovirus-2 (PCV-2) is very important for surveillance of postweaning multisystemic wasting syndrome. Droplet digital polymerase chain reaction (ddPCR) is novel PCR method that can achieve high precision. Our study aimed to develop a sensitive assay utilizing ddPCR to detect PCV-2. Specificity of the assay was confirmed by the failure of amplification of DNA of other relevant viruses. The detection limit for ddPCR was 25 copies/μL, a 4-fold greater sensitivity than TaqMan real-time PCR. Both methods showed a high degree of linearity (R(2) = ~1), although TaqMan real-time PCR showed less sensitivity than ddPCR for clinical detection. Our findings indicate that ddPCR might represent a promising platform for detecting PCV-2 viral loads.
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Zhao et al. (2015) studied Porcine circovirus-2 (PCV-2) infection. Droplet digital polymerase chain reaction (ddPCR) vs. TaqMan real-time PCR was evaluated on Detection limit for PCV-2 (4-fold greater sensitivity). Droplet digital PCR detected Porcine circovirus-2 with a detection limit of 25 copies/μL, demonstrating a 4-fold greater sensitivity than TaqMan real-time PCR.
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