PMID- 23850717
OWN - NLM
STAT- MEDLINE
DCOM- 20140418
LR  - 20191210
IS  - 1879-0984 (Electronic)
IS  - 0166-0934 (Linking)
VI  - 193
IP  - 2
DP  - 2013 Nov
TI  - Development of a loop-mediated isothermal amplification assay for the detection 
      of porcine hokovirus.
PG  - 415-8
LID - S0166-0934(13)00257-7 [pii]
LID - 10.1016/j.jviromet.2013.06.040 [doi]
AB  - Hokoviruses have recently been detected as pathogens belonging to the family 
      Parvoviridae, which comprises porcine hokovirus (PHoV) and bovine hokovirus 
      (BHoV). In this study, we developed a loop-mediated isothermal amplification 
      (LAMP) assay for the rapid, specific and sensitive detection of PHoV. A set of 
      four primers specific for six regions within the PHoV VP1/2 genes was designed 
      using online software. The reaction temperature and time were optimized at 65 degrees C 
      and 60 min, respectively. LAMP products were detected by agarose gel 
      electrophoresis or by visual inspection of a color change caused by a fluorescent 
      dye. The method was highly specific for PHoV, and no cross-reaction was observed 
      with porcine circovirus type 2 (PCV2), porcine parvovirus (PPV), porcine 
      bocavirus (PBoV), pseudorabies virus (PRV), porcine reproductive and respiratory 
      syndrome virus (PRRSV), classic swine fever virus (CSFV), or Japanese 
      encephalitis virus (JEV). The detection limit was approximately 10 copies per 
      reaction, which was 10 times more sensitive than conventional PCR. Furthermore, 
      the efficiency of detection of PHoV in clinical samples was comparable to that of 
      PCR and sequencing. These results show that the LAMP assay is a simple, rapid, 
      sensitive and specific method for detecting PHoV. It does not require specialized 
      equipment and can be used to detect PHoV both in the laboratory and in the field.
CI  - Copyright (c) 2013 Elsevier B.V. All rights reserved.
FAU - Li, Bin
AU  - Li B
AD  - Institute of Veterinary Medicine, Jiangsu Academy of Agricultural Sciences; Key 
      Laboratory of Veterinary Biological Engineering and Technology, Ministry of 
      Agriculture; National Center for Engineering Research of Veterinary Bio-products, 
      Nanjing 210014, Jiangsu Province, China. Electronic address: libinana@126.com.
FAU - Sun, Bing
AU  - Sun B
FAU - Du, Lu-ping
AU  - Du LP
FAU - Mao, Ai-hua
AU  - Mao AH
FAU - Wen, Li-bin
AU  - Wen LB
FAU - Ni, Yan-xiu
AU  - Ni YX
FAU - Zhang, Xue-han
AU  - Zhang XH
FAU - He, Kong-wang
AU  - He KW
LA  - eng
PT  - Evaluation Study
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20130709
PL  - Netherlands
TA  - J Virol Methods
JT  - Journal of virological methods
JID - 8005839
RN  - 0 (DNA Primers)
RN  - 0 (Viral Structural Proteins)
SB  - IM
MH  - Animals
MH  - DNA Primers/genetics
MH  - Molecular Diagnostic Techniques/*methods
MH  - Nucleic Acid Amplification Techniques/*methods
MH  - Parvoviridae Infections/diagnosis/*veterinary
MH  - Parvovirinae/*isolation & purification
MH  - Sensitivity and Specificity
MH  - Swine
MH  - Swine Diseases/*diagnosis/virology
MH  - Temperature
MH  - Time Factors
MH  - Veterinary Medicine/*methods
MH  - Viral Structural Proteins/genetics
OTO - NOTNLM
OT  - Detection
OT  - Loop-mediated isothermal amplification
OT  - PCR
OT  - Porcine hokovirus
EDAT- 2013/07/16 06:00
MHDA- 2014/04/20 06:00
CRDT- 2013/07/16 06:00
PHST- 2013/03/13 00:00 [received]
PHST- 2013/06/18 00:00 [revised]
PHST- 2013/06/26 00:00 [accepted]
PHST- 2013/07/16 06:00 [entrez]
PHST- 2013/07/16 06:00 [pubmed]
PHST- 2014/04/20 06:00 [medline]
AID - S0166-0934(13)00257-7 [pii]
AID - 10.1016/j.jviromet.2013.06.040 [doi]
PST - ppublish
SO  - J Virol Methods. 2013 Nov;193(2):415-8. doi: 10.1016/j.jviromet.2013.06.040. Epub 
      2013 Jul 9.