Sensitive detection of Porcine circovirus-2 by droplet digital polymerase chain reaction
| Autor: | Shan Zhao, Lixin Xi, Yan Yubao, Lin Hua, Qigui Yan, Chen Shijie, Zhongxiang Hao, Chen Zhenrong, Hu Juan, Sun Yingjie, Miao Yang, Caifang Wen |
|---|---|
| Rok vydání: | 2015 |
| Předmět: |
Circovirus
Swine Diseases Detection limit China General Veterinary Swine Viral Load Biology Real-Time Polymerase Chain Reaction biology.organism_classification Sensitivity and Specificity Virology Molecular biology Porcine circovirus Real-time polymerase chain reaction Limit of Detection DNA Viral TaqMan Animals Digital polymerase chain reaction Pcr method Circoviridae Infections Dna viral |
| Zdroj: | Journal of Veterinary Diagnostic Investigation. 27:784-788 |
| ISSN: | 1943-4936 1040-6387 |
| DOI: | 10.1177/1040638715608358 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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