PMID- 32456688
OWN - NLM
STAT- MEDLINE
DCOM- 20210104
LR  - 20210104
IS  - 1746-6148 (Electronic)
IS  - 1746-6148 (Linking)
VI  - 16
IP  - 1
DP  - 2020 May 26
TI  - A novel protein chip for simultaneous detection of antibodies against four 
      epidemic swine viruses in China.
PG  - 162
LID - 10.1186/s12917-020-02375-7 [doi]
LID - 162
AB  - BACKGROUND: At present, pig industry in China is faced with the complex situation 
      of mixed infection caused by multiple pathogens. It is urgent to develop some new 
      high-throughput molecular diagnosis assays to simultaneously detect pathogens or 
      antibodies. Biochip array technology has made it possible to screen thousands of 
      samples simultaneously; it has been twice named as one of the top 10 scientific 
      and technological breakthroughs. Studies have reported encouraging results using 
      protein biochips for detecting antibodies against avian infectious bronchitis 
      virus and ruminant bluetongue virus, but the research of this technology for the 
      diagnosis of swine diseases is still sparse. RESULTS: In this study, a novel 
      protein chip was developed that can simultaneously detect the antibodies of four 
      important swine viruses as follow, classical swine fever virus (CSFV), porcine 
      parvovirus (PPV), Japanese encephalitis virus (JEV), and porcine reproductive and 
      respiratory syndrome virus (PRRSV). Four prokaryotic expression plasmids 
      pET-32a-E2 of CSFV, -VP2 of PPV, -EDIII of JEV, and -N of PRRSV were induced by 
      IPTG (Isopropyl beta-D-1-Thiogalactopyranoside) and overexpressed in E.coli, 
      respectively. The purified proteins were identified by Western blotting and then 
      printed on epoxy-coated glass slides. The optimized parameters of this diagnostic 
      chip showed that the spotting concentrations of E2、VP2、EDIII、N proteins were 0.2, 
      0.4, 0.4, and 0.4 mg/mL. The optimal primary and secondary antibody dilutions 
      were 1:50 and 1: 600. Compared with the commercial ELISA (Enzyme-linked 
      immunosorbent assay) kits, the positive and negative coincidence rates of this 
      chip were 95.8% ~ 100 and 86.2% ~ 100%, as well as, no cross-reaction. 
      CONCLUSION: This protein chip provided a fast, specific, and sensitive method for 
      simultaneous detection of antibodies in clinical serum samples. Compared with 
      traditional methods, this protein chip can monitor very small amount of serum.
FAU - Wu, Yue
AU  - Wu Y
AD  - MOE Joint International Research Laboratory of Animal Health and Food Safety, 
      College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, 210095, 
      China.
FAU - Wu, Xudan
AU  - Wu X
AD  - MOE Joint International Research Laboratory of Animal Health and Food Safety, 
      College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, 210095, 
      China.
FAU - Chen, Jing
AU  - Chen J
AD  - MOE Joint International Research Laboratory of Animal Health and Food Safety, 
      College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, 210095, 
      China.
FAU - Hu, Jingfei
AU  - Hu J
AD  - College of Veterinary Medicine, Sichuan Agricultural University, Chengdu, 611130, 
      China.
FAU - Huang, Xiaobo
AU  - Huang X
AD  - College of Veterinary Medicine, Sichuan Agricultural University, Chengdu, 611130, 
      China.
FAU - Zhou, Bin
AU  - Zhou B
AUID- ORCID: 0000-0001-7279-9489
AD  - MOE Joint International Research Laboratory of Animal Health and Food Safety, 
      College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, 210095, 
      China. zhoubin@njau.edu.cn.
LA  - eng
GR  - 2016YFD0500701-3/the National Key Research and Development Program/
PT  - Journal Article
DEP - 20200526
PL  - England
TA  - BMC Vet Res
JT  - BMC veterinary research
JID - 101249759
RN  - 0 (Antibodies, Viral)
SB  - IM
MH  - Animals
MH  - Antibodies, Viral/*blood/immunology
MH  - Classical Swine Fever Virus/immunology
MH  - Encephalitis Virus, Japanese/immunology
MH  - Parvovirus, Porcine/immunology
MH  - Porcine respiratory and reproductive syndrome virus/immunology
MH  - Protein Array Analysis/methods/*veterinary
MH  - Swine
MH  - Swine Diseases/*diagnosis/immunology/*virology
PMC - PMC7249397
OTO - NOTNLM
OT  - Protein biochip
OT  - Simultaneous detection
OT  - Swine diseases, antibodies
COIS- The authors declare that they have no competing interests.
EDAT- 2020/05/28 06:00
MHDA- 2021/01/05 06:00
PMCR- 2020/05/26
CRDT- 2020/05/28 06:00
PHST- 2020/02/13 00:00 [received]
PHST- 2020/05/14 00:00 [accepted]
PHST- 2020/05/28 06:00 [entrez]
PHST- 2020/05/28 06:00 [pubmed]
PHST- 2021/01/05 06:00 [medline]
PHST- 2020/05/26 00:00 [pmc-release]
AID - 10.1186/s12917-020-02375-7 [pii]
AID - 2375 [pii]
AID - 10.1186/s12917-020-02375-7 [doi]
PST - epublish
SO  - BMC Vet Res. 2020 May 26;16(1):162. doi: 10.1186/s12917-020-02375-7.