PMID- 25241142
OWN - NLM
STAT- MEDLINE
DCOM- 20151012
LR  - 20191210
IS  - 1879-0984 (Electronic)
IS  - 0166-0934 (Linking)
VI  - 209
DP  - 2014 Dec
TI  - Rapid and sensitive detection of type II porcine reproductive and respiratory 
      syndrome virus by reverse transcription loop-mediated isothermal amplification 
      combined with a vertical flow visualization strip.
PG  - 86-94
LID - S0166-0934(14)00353-X [pii]
LID - 10.1016/j.jviromet.2014.09.011 [doi]
AB  - Reverse transcription-loop-mediated isothermal amplification (RT-LAMP) was 
      combined with a vertical flow (VF) nucleic acid detection strip to develop a 
      universal assay for the detection of type II porcine reproductive and respiratory 
      syndrome virus (PRRSV). The loop primers were labeled separately with biotin and 
      fluorescein isothiocyanate (FITC) in this assay. Using optimized parameters, the 
      whole reaction could be completed in <50 min in a completely enclosed 
      environment. The detection limit of this assay was found to be 1 pg RNA, 30 
      tissue culture infective dose 50 (TCID50) virus, or 230 copies of recombinant 
      plasmid DNA, which is relatively higher than that of RT-LAMP analyzed by agarose 
      gel, RT-LAMP visualized by calcein, and the conventional RT-polymerase chain 
      reaction (PCR). No false-positive results were obtained in the specificity assay. 
      The efficiency of the RT-LAMP method was tested by analyzing 43 clinical samples, 
      and the results were compared with those obtained by RT-PCR analysis, with the 
      respective positive rates of 32.56% and 27.91%. This result confirmed that the 
      method described is a rapid, accurate, and sensitive method for universal type II 
      PRRSV detection. Also, this method can be used for the rapid detection of type II 
      PRRSV during the early phase of an outbreak, especially for rapid veterinary 
      diagnosis on the spot and in rural areas.
CI  - Copyright (c) 2014 Elsevier B.V. All rights reserved.
FAU - Gou, Hongchao
AU  - Gou H
AD  - College of Veterinary Medicine, South China Agricultural University, Guangzhou, 
      China.
FAU - Deng, Jieru
AU  - Deng J
AD  - College of Veterinary Medicine, South China Agricultural University, Guangzhou, 
      China.
FAU - Pei, Jingjing
AU  - Pei J
AD  - College of Veterinary Medicine, South China Agricultural University, Guangzhou, 
      China.
FAU - Wang, Jiaying
AU  - Wang J
AD  - College of Veterinary Medicine, South China Agricultural University, Guangzhou, 
      China.
FAU - Liu, Wenjun
AU  - Liu W
AD  - College of Veterinary Medicine, South China Agricultural University, Guangzhou, 
      China.
FAU - Zhao, Mingqiu
AU  - Zhao M
AD  - College of Veterinary Medicine, South China Agricultural University, Guangzhou, 
      China.
FAU - Chen, Jinding
AU  - Chen J
AD  - College of Veterinary Medicine, South China Agricultural University, Guangzhou, 
      China. Electronic address: jdchen@scau.edu.cn.
LA  - eng
PT  - Evaluation Study
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20140918
PL  - Netherlands
TA  - J Virol Methods
JT  - Journal of virological methods
JID - 8005839
RN  - 0 (DNA Primers)
SB  - IM
MH  - Animals
MH  - Chromatography/*methods
MH  - DNA Primers/genetics
MH  - False Positive Reactions
MH  - Molecular Diagnostic Techniques/*methods
MH  - Nucleic Acid Amplification Techniques/*methods
MH  - Porcine Reproductive and Respiratory Syndrome/*diagnosis/virology
MH  - Porcine respiratory and reproductive syndrome virus/*isolation & purification
MH  - Reverse Transcription
MH  - Sensitivity and Specificity
MH  - Swine
MH  - Time Factors
MH  - Veterinary Medicine/*methods
OTO - NOTNLM
OT  - Loop-mediated isothermal amplification
OT  - Porcine reproductive and respiratory syndrome virus
OT  - Vertical flow visualization strip
EDAT- 2014/09/23 06:00
MHDA- 2015/10/13 06:00
CRDT- 2014/09/22 06:00
PHST- 2014/02/19 00:00 [received]
PHST- 2014/08/30 00:00 [revised]
PHST- 2014/09/09 00:00 [accepted]
PHST- 2014/09/22 06:00 [entrez]
PHST- 2014/09/23 06:00 [pubmed]
PHST- 2015/10/13 06:00 [medline]
AID - S0166-0934(14)00353-X [pii]
AID - 10.1016/j.jviromet.2014.09.011 [doi]
PST - ppublish
SO  - J Virol Methods. 2014 Dec;209:86-94. doi: 10.1016/j.jviromet.2014.09.011. Epub 
      2014 Sep 18.