Sensitive detection of Porcine circovirus-2 by droplet digital polymerase chain reaction

• Published in: Journal of veterinary diagnostic investigation 2015-11, Vol.27 (6), p.784-788
• Main Authors: Zhao, Shan, Lin, Hua, Chen, Shijie, Yang, Miao, Yan, Qigui, Wen, Caifang, Hao, Zhongxiang, Yan, Yubao, Sun, Yingjie, Hu, Juan, Chen, Zhenrong, Xi, Lixin
• Format: Article
• Language: English
• Subjects: Animals; China; Circoviridae Infections - diagnosis; Circoviridae Infections - veterinary; Circoviridae Infections - virology; Circovirus - genetics; Circovirus - isolation & purification; DNA, Viral - analysis; Limit of Detection; Real-Time Polymerase Chain Reaction - methods; Real-Time Polymerase Chain Reaction - veterinary; Sensitivity and Specificity; Swine; Swine Diseases - diagnosis; Swine Diseases - virology; Viral Load - veterinary
• ISSN: 1040-6387; 1943-4936
• DOI: 10.1177/1040638715608358

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 (R2 = ~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.