Rapid and specific detection of porcine parvovirus using real-time PCR and High Resolution Melting (HRM) analysis

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• 作者：Hai-Qiong Yu (1)
  Xian-Quan Cai (2)
  Zhi-Xiong Lin (1)
  Xiang-Li Li (3)
  Qiao-Yun Yue (2)
  Rong Li (2)
  Xing-Quan Zhu (4)
  
  1. Technical Center ; Guangdong Entry-Exit Inspection and Quarantine Bureau ; Guangzhou ; Guangdong Province ; 510630 ; PR China
  2. Technical Center ; Zhongshan Entry-Exit Inspection and Quarantine Bureau ; Zhongshan ; Guangdong Province ; 528403 ; PR China
  3. Zhongshan Torch Polytechnic College ; Zhongshan ; Guangdong Province ; 528403 ; PR China
  4. State Key Laboratory of Veterinary Etiological Biology ; Lanzhou Veterinary Research Institute ; Chinese Academy of Agricultural Sciences ; Lanzhou ; Gansu Province ; 730046 ; PR China
  
• 关键词：Porcine parvovirus (PPV) ; High resolution melting (HRM) ; Real ; time PCR
• 刊名：BMC Veterinary Research
• 出版年：2015
• 出版时间：December 2015
• 年：2015
• 卷：11
• 期：1
• 全文大小：684 KB
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• 刊物主题：Veterinary Medicine; Zoology; Transgenics;
• 出版者：BioMed Central
• ISSN：1746-6148

文摘

Background Porcine parvovirus (PPV) is the important causative agent for infectious infertility, which is a fairly tough virus that multiplies normally in the intestine of pigs without causing clinical signs in the world. Results We developed an assay integrating real-time PCR and high resolution melting (HRM) analysis for the detection of PPV. Primers targeting the VP gene were highly specific, as evidenced by the negative amplification of closely related viruses, such as porcine circovirus 2 (PCV2), porcine reproductive and respiratory syndrome virus (PRRSV), pseudorabies virus (PRV), classical swine fever virus (CSFV), or Japanese encephalitis virus (JEV). The performance of unlabeled real time PCR was compared to TaqMan real time PCR, and the detection limits of the two methods were nearly equal. Moreover, there was good correlation between Cp and diluted genomic DNA when tested with the two methods. The assay has the accuracy of 100% in reference to labeled real time PCR, when it was tested on 45 clinical samples. Conclusions The present study demonstrated that the established assay integrating real-time PCR and HRM is relatively cost-effective and more stable, which provides an alternative tool for rapid, simple, specific and sensitive detection of PPV.