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Development and clinical validation of a Potential Penside Colorimetric LAMP Assay of Porcine Circovirus type 3
  • +12
  • Yang Wang,
  • Jie Zhang,
  • Miaomiao Li,
  • Yunwen Ou,
  • Danian Chen,
  • Yaozhong Ding,
  • Weibing Zhang,
  • Yanjun Li,
  • Qian Hou,
  • Xiaoyun Li,
  • Luoyi Zhou,
  • Kataryna Podgorska,
  • Alexei Zaberezhny,
  • Anna Szczotka-Bochniarz,
  • Yong-sheng Liu
Yang Wang
State Key Laboratory of Veterinary Etiological Biology

Corresponding Author:[email protected]

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Jie Zhang
Hebei Normal University of Science and Technology
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Miaomiao Li
State Key Laboratory of Veterinary Etiological Biology
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Yunwen Ou
Animal Disease Prevention and Control Center of Kaijiang County China
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Danian Chen
State Key Laboratory of Veterinary Etiological Biology
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Yaozhong Ding
State Key Laboratory of Veterinary Etiological Biology
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Weibing Zhang
State Key Laboratory of Veterinary Etiological Biology
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Yanjun Li
State Key Laboratory of Veterinary Etiological Biology
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Qian Hou
State Key Laboratory of Veterinary Etiological Biology
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Xiaoyun Li
State Key Laboratory of Veterinary Etiological Biology
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Luoyi Zhou
Hebei Normal University of Science and Technology
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Kataryna Podgorska
Department of Swine Diseases National Veterinary Research Institute 57 Partyzantow 24-100 Puławy Poland
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Alexei Zaberezhny
Federal State Budgetary Institution All-Russian Research and Technological Institute of Biological Industry (VNITIBP) Moscow Russia
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Anna Szczotka-Bochniarz
Department of Swine Diseases National Veterinary Research Institute 57 Partyzantow 24-100 Puławy Poland
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Yong-sheng Liu
Hebei Normal University of Science and Technology
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Abstract

OBJECTIVES: Porcine circovirus type 3 (PCV3), a novel circovirus, imposes great burdens on the global pig industry. The penside tests for detecting PCV3 are critical for assessing the epidemiological status and adjusting disease prevention in the context of no vaccine available. METHODS: A one-step molecular assay based on visual loop-mediated isothermal amplification (vLAMP) was developed for simple and rapid detection of PCV3. We compared its sensitivity and specificity with TaqMan quantitative real-time polymerase chain reaction (qPCR) and applied the developed assay in the epidemiological study of 407 pooled swine sera collected in the whole China mainland during 2017-2018. We also explored the feasibility of the vLAMP assay for detecting crude samples without a prior DNA isolation step to expand its application field. RESULTS: Results showed that the vLAMP assay reliably detected PCV3 cap with a equal 10 DNA copies detection limit compared to that of Taqman qPCR assay. In the epidemiological study, the PCV3 positive detection rate for 407 swine pooled sera detected by the vLAMP assay was 37.35% (152/407) while it was 39.01% (159/407) for Taqman qPCR. For the direct-detection method, the results kept perfect specificity (100%) but displayed lower sensitivity (100% for CT<32), indicating the method not suited discrimination of low viral loads samples. CONCLUSIONS: The one-step vLAMP is a convenient, rapid, and cost-effective diagnostic validation procedure on penside and will enable epidemiological surveillance of PCV3 which has widely spread in China mainland.