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Development of Isothermal Recombinase Polymerase Amplification Assay for Rapid Detection of Porcine Circovirus Type 2.

Abstract Porcine circovirus virus type II (PCV2) is the etiology of postweaning multisystemic wasting syndrome (PMWS), porcine dermatitis, nephropathy syndrome (PDNS), and necrotizing pneumonia. Rapid diagnosis tool for detection of PCV2 plays an important role in the disease control and eradication program. Recombinase polymerase amplification (RPA) assays using a real-time fluorescent detection (PCV2 real-time RPA assay) and RPA combined with lateral flow dipstick (PCV2 RPA LFD assay) were developed targeting the PCV2 ORF2 gene. The results showed that the sensitivity of the PCV2 real-time RPA assay was 10(2) copies per reaction within 20 min at 37°C and the PCV2 RPA LFD assay had a detection limit of 10(2) copies per reaction in less than 20 min at 37°C. Both assays were highly specific for PCV2, with no cross-reactions with porcine circovirus virus type 1, foot-and-mouth disease virus, pseudorabies virus, porcine parvovirus, porcine reproductive and respiratory syndrome virus, and classical swine fever virus. Therefore, the RPA assays provide a novel alternative for simple, sensitive, and specific identification of PCV2.
PMID
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Authors

Mayor MeshTerms
Keywords
Journal Title biomed research international
Publication Year Start




PMID- 28424790
OWN - NLM
STAT- In-Process
DA  - 20170420
LR  - 20170423
IS  - 2314-6141 (Electronic)
VI  - 2017
DP  - 2017
TI  - Development of Isothermal Recombinase Polymerase Amplification Assay for Rapid
      Detection of Porcine Circovirus Type 2.
PG  - 8403642
LID - 10.1155/2017/8403642 [doi]
AB  - Porcine circovirus virus type II (PCV2) is the etiology of postweaning
      multisystemic wasting syndrome (PMWS), porcine dermatitis, nephropathy syndrome
      (PDNS), and necrotizing pneumonia. Rapid diagnosis tool for detection of PCV2
      plays an important role in the disease control and eradication program.
      Recombinase polymerase amplification (RPA) assays using a real-time fluorescent
      detection (PCV2 real-time RPA assay) and RPA combined with lateral flow dipstick 
      (PCV2 RPA LFD assay) were developed targeting the PCV2 ORF2 gene. The results
      showed that the sensitivity of the PCV2 real-time RPA assay was 102 copies per
      reaction within 20 min at 37 degrees C and the PCV2 RPA LFD assay had a detection
      limit of 102 copies per reaction in less than 20 min at 37 degrees C. Both assays
      were highly specific for PCV2, with no cross-reactions with porcine circovirus
      virus type 1, foot-and-mouth disease virus, pseudorabies virus, porcine
      parvovirus, porcine reproductive and respiratory syndrome virus, and classical
      swine fever virus. Therefore, the RPA assays provide a novel alternative for
      simple, sensitive, and specific identification of PCV2.
FAU - Yang, Yang
AU  - Yang Y
AD  - State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary
      Research Institute, Chinese Academy of Agricultural Sciences, Xujiaping 1,
      Lanzhou, Gansu 730046, China.
FAU - Qin, Xiaodong
AU  - Qin X
AD  - State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary
      Research Institute, Chinese Academy of Agricultural Sciences, Xujiaping 1,
      Lanzhou, Gansu 730046, China.
FAU - Sun, Yingjun
AU  - Sun Y
AD  - State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary
      Research Institute, Chinese Academy of Agricultural Sciences, Xujiaping 1,
      Lanzhou, Gansu 730046, China.
FAU - Cong, Guozheng
AU  - Cong G
AD  - State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary
      Research Institute, Chinese Academy of Agricultural Sciences, Xujiaping 1,
      Lanzhou, Gansu 730046, China.
FAU - Li, Yanmin
AU  - Li Y
AD  - State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary
      Research Institute, Chinese Academy of Agricultural Sciences, Xujiaping 1,
      Lanzhou, Gansu 730046, China.
FAU - Zhang, Zhidong
AU  - Zhang Z
AUID- ORCID: 0000-0002-2186-7661
AD  - State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary
      Research Institute, Chinese Academy of Agricultural Sciences, Xujiaping 1,
      Lanzhou, Gansu 730046, China.
LA  - eng
PT  - Journal Article
DEP - 20170323
PL  - United States
TA  - Biomed Res Int
JT  - BioMed research international
JID - 101600173
PMC - PMC5382309
EDAT- 2017/04/21 06:00
MHDA- 2017/04/21 06:00
CRDT- 2017/04/21 06:00
PHST- 2016/12/06 [received]
PHST- 2017/03/13 [accepted]
AID - 10.1155/2017/8403642 [doi]
PST - ppublish
SO  - Biomed Res Int. 2017;2017:8403642. doi: 10.1155/2017/8403642. Epub 2017 Mar 23.

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