PMID- 31351984
OWN - NLM
STAT- MEDLINE
DCOM- 20200504
LR  - 20200505
IS  - 1879-0984 (Electronic)
IS  - 0166-0934 (Linking)
VI  - 272
DP  - 2019 Oct
TI  - Fluorescence resonance energy transfer combined with asymmetric PCR for broad and 
      sensitive detection of porcine reproductive and respiratory syndrome virus 2.
PG  - 113710
LID - S0166-0934(18)30456-7 [pii]
LID - 10.1016/j.jviromet.2019.113710 [doi]
AB  - With its ever-increasing viral genetic diversity, accurate diagnosis of porcine 
      reproductive and respiratory syndrome virus (PRRSV) infection is indispensable 
      for PRRSV control. Here, a sensitive graphene oxide (GO)-based FRET method was 
      developed to detect PRRSV-2 based on the ability of GO to quench fluorophore by 
      fluorescence resonance energy transfer (FRET). Using primers and a 
      fluorophore-labeled ssDNA probe targeting a conserved region between the PRRSV M 
      gene and 3'UTR, asymmetric PCR specifically amplified viral ssDNA that could 
      anneal with probe to generate dsDNA only in the presence of virus. Upon 
      exonuclease III treatment to release the probe fluorophore, which degrades dsDNA 
      with blunt ends or recessed 3 -termini, the ssDNA annealed with other probe to 
      generate enhanced fluorescence. This GO-based FRET assay specifically detected 
      both classical and highly pathogenic PRRSV, with analytical sensitivity 
      approaching 10 copies/muL, similar to that of real-time PCR but greater than that 
      of conventional reverse transcription PCR (RT-PCR). Consistent with real-time 
      RT-PCR detection, the assay developed here exhibited high diagnostic sensitivity 
      for virus detection of sera from experimentally and naturally infected pigs. 
      Thus, this novel GO-based FRET assay combined with asymmetric PCR detection is 
      sensitive and specific and will be valuable for future PRRSV diagnosis.
CI  - Copyright (c) 2019 Elsevier B.V. All rights reserved.
FAU - Guo, Wenping
AU  - Guo W
AD  - Department of Preventive Veterinary Medicine, College of Veterinary Medicine, 
      Northwest A&F University, Scientific Observing and Experimental Station of 
      Veterinary Pharmacology and Diagnostic Technology, Ministry of Agriculture, 
      Yangling, 712100, China.
FAU - Xu, Gang
AU  - Xu G
AD  - Department of Preventive Veterinary Medicine, College of Veterinary Medicine, 
      Northwest A&F University, Scientific Observing and Experimental Station of 
      Veterinary Pharmacology and Diagnostic Technology, Ministry of Agriculture, 
      Yangling, 712100, China.
FAU - Wang, Yihan
AU  - Wang Y
AD  - Department of Preventive Veterinary Medicine, College of Veterinary Medicine, 
      Northwest A&F University, Scientific Observing and Experimental Station of 
      Veterinary Pharmacology and Diagnostic Technology, Ministry of Agriculture, 
      Yangling, 712100, China.
FAU - Nan, Yuchen
AU  - Nan Y
AD  - Department of Preventive Veterinary Medicine, College of Veterinary Medicine, 
      Northwest A&F University, Scientific Observing and Experimental Station of 
      Veterinary Pharmacology and Diagnostic Technology, Ministry of Agriculture, 
      Yangling, 712100, China.
FAU - Wu, Chunyan
AU  - Wu C
AD  - Department of Preventive Veterinary Medicine, College of Veterinary Medicine, 
      Northwest A&F University, Scientific Observing and Experimental Station of 
      Veterinary Pharmacology and Diagnostic Technology, Ministry of Agriculture, 
      Yangling, 712100, China.
FAU - Zhou, En-Min
AU  - Zhou EM
AD  - Department of Preventive Veterinary Medicine, College of Veterinary Medicine, 
      Northwest A&F University, Scientific Observing and Experimental Station of 
      Veterinary Pharmacology and Diagnostic Technology, Ministry of Agriculture, 
      Yangling, 712100, China. Electronic address: zhouem@nwsuaf.edu.cn.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20190725
PL  - Netherlands
TA  - J Virol Methods
JT  - Journal of virological methods
JID - 8005839
RN  - 0 (DNA Primers)
RN  - 0 (DNA Probes)
RN  - 0 (RNA, Viral)
RN  - 0 (graphene oxide)
RN  - 7782-42-5 (Graphite)
SB  - IM
MH  - Animals
MH  - Base Sequence
MH  - Cell Line
MH  - DNA Primers/genetics
MH  - DNA Probes
MH  - Fluorescence Resonance Energy Transfer/*methods/*veterinary
MH  - Graphite
MH  - Polymerase Chain Reaction/*methods/*veterinary
MH  - Porcine Reproductive and Respiratory Syndrome/blood/diagnosis/virology
MH  - Porcine respiratory and reproductive syndrome virus/*genetics/*isolation & 
      purification
MH  - RNA, Viral/genetics
MH  - Reverse Transcriptase Polymerase Chain Reaction/methods/veterinary
MH  - Sensitivity and Specificity
MH  - Swine
MH  - Swine Diseases/blood/diagnosis/virology
OTO - NOTNLM
OT  - Asymmetric PCR
OT  - FRET
OT  - Graphene oxide
OT  - PRRSV
EDAT- 2019/07/29 06:00
MHDA- 2020/05/06 06:00
CRDT- 2019/07/29 06:00
PHST- 2018/09/14 00:00 [received]
PHST- 2019/07/24 00:00 [revised]
PHST- 2019/07/24 00:00 [accepted]
PHST- 2019/07/29 06:00 [pubmed]
PHST- 2020/05/06 06:00 [medline]
PHST- 2019/07/29 06:00 [entrez]
AID - S0166-0934(18)30456-7 [pii]
AID - 10.1016/j.jviromet.2019.113710 [doi]
PST - ppublish
SO  - J Virol Methods. 2019 Oct;272:113710. doi: 10.1016/j.jviromet.2019.113710. Epub 
      2019 Jul 25.