Rapid detection of porcine circovirus type 2 using a TaqMan-based real-time PCR

• Main Authors: Zhang Chunling, Xu Yan, Li Chunhua, Zhu Lianlong, Zou Yong, Han Fangting, Zhao Kai, Tan Furong, Wang Jinbin, Tao Shiru, He Xizhong, Zhou Zongqing, Tang Xueming
• Format: Article
• Language: English
• Published: BMC 2010-12-01
• Series: Virology Journal
• Online Access: http://www.virologyj.com/content/7/1/374

<p>Abstract</p> <p>Porcine circovirus type 2 (PCV2) and the associated disease postweaning multisystemic wasting syndrome (PMWS) have caused heavy losses in global agriculture in recent decades. Rapid detection of PCV2 is very important for the effective prophylaxis and treatment of PMWS. To establish a sensitive, specific assay for the detection and quantitation of PCV2, we designed and synthesized specific primers and a probe in the open reading frame 2. The assay had a wide dynamic range with excellent linearity and reliable reproducibility, and detected between 10²and 10¹⁰copies of the genomic DNA per reaction. The coefficient of variation for Ct values varied from 0.59% to 1.05% in the same assay and from 1.9% to 4.2% in 10 different assays. The assay did not cross-react with porcine circovirus type 1, porcine reproductive and respiratory, porcine epidemic diarrhea, transmissible gastroenteritis of pigs and rotavirus. The limits of detection and quantitation were 10 and 100 copies, respectively. Using the established real-time PCR system, 39 of the 40 samples we tested were detected as positive.</p>