Novel Approach for Isolation and Identification of Porcine Epidemic Diarrhea Virus (PEDV) Strain NJ Using Porcine Intestinal Epithelial Cells
Abstract
:1. Introduction
2. Materials and Methods
2.1. Cells and Clinical Samples
2.2. RNA Extraction and RT-PCR Assay
2.3. Virus Isolation
2.4. Electron Microscopy Assay
2.5. Immunofluorescence Assay
2.6. Sequence Alignment and Phylogenetic Analysis of the S Gene
2.7. Virulence Experiment for IEC-Adapted NJ
3. Results
3.1. Virus Isolation
3.2. Determination of Viral Titer
3.3. Electron Microscopy
3.4. Immunofluorescence
3.5. Phylogenetic Analysis of the S Gene
3.6. Amino Acid Variability of the S Protein of IEC-Adapted NJ after Serial Passaging
3.7. Pathogenicity Analysis of PEDV Strain NJ
4. Discussion
Acknowledgments
Author Contributions
Conflicts of Interest
References
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Strain | ID | Country | Strain | ID | Country |
---|---|---|---|---|---|
83P-5 (parent) | AB548618 | Japan | HLJ-2012 | JX512907 | China |
83P-5 (34th) | AB548619 | Japan | IA1 | KF468753 | USA |
83P-5 (100th) | AB548621 | Japan | IA2 | KF468754 | USA |
AD02 | KC879281 | South Korea | ISU13-19338E-IN-homogenate | KF650370 | USA |
AH2012 | KC210145 | China | ISU13-19338E-IN-passage3 | KF650371 | USA |
BJ-2012-1 | JX435299 | China | ISU13-19338E-IN-passage9 | KF650372 | USA |
Br1/87 | Z25483 | France | JS2008 | KC109141 | China |
CH9-FJ | JQ979287 | China | JS-HZ2012 | KC210147 | China |
CH22-JS | JQ979290 | China | KH | AB548622 | Japan |
CHGD-01 | JN980698 | China | LJB/03 | DQ985739 | China |
Chinju99 | AY167585 | Korea | MK | AB548624 | Japan |
CH/S | JN547228 | China | MN | KF468752 | USA |
CV777 vaccine | JN599150 | China | NK | AB548623 | Japan |
CV777 | AF353511 | Belgium | YN1 | KT021227 | China |
DR13 | JQ023161 | Korea | YN15 | KT021228 | China |
DR13 (Attenuated) | JQ023162 | Korea | YN144 | KT021232 | China |
DX | JN104080 | China |
Strain | Amino Acid Position | ||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|
3 | 15 | 70 | 114 | 282 | 324 | 378 | 438 | 973 | 1023 | 1167 | |
NJ(Parent) | S | S | A | N | L | S | N | I | Y | K | A |
NJ(15th) | S | S | D | N | L | S | N | I | H | N | D |
NJ(30th) | A | S | D | S | L | S | K | I | H | N | D |
NJ(45th) | A | L | D | S | W | R | K | L | H | N | D |
CV777 | S | P | A | N | L | S | N | I | Y | K | A |
CV777 vaccine | P | L | D | S | W | F | K | V | H | N | D |
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Shi, W.; Jia, S.; Zhao, H.; Yin, J.; Wang, X.; Yu, M.; Ma, S.; Wu, Y.; Chen, Y.; Fan, W.; et al. Novel Approach for Isolation and Identification of Porcine Epidemic Diarrhea Virus (PEDV) Strain NJ Using Porcine Intestinal Epithelial Cells. Viruses 2017, 9, 19. https://doi.org/10.3390/v9010019
Shi W, Jia S, Zhao H, Yin J, Wang X, Yu M, Ma S, Wu Y, Chen Y, Fan W, et al. Novel Approach for Isolation and Identification of Porcine Epidemic Diarrhea Virus (PEDV) Strain NJ Using Porcine Intestinal Epithelial Cells. Viruses. 2017; 9(1):19. https://doi.org/10.3390/v9010019
Chicago/Turabian StyleShi, Wen, Shuo Jia, Haiyuan Zhao, Jiyuan Yin, Xiaona Wang, Meiling Yu, Sunting Ma, Yang Wu, Ying Chen, Wenlu Fan, and et al. 2017. "Novel Approach for Isolation and Identification of Porcine Epidemic Diarrhea Virus (PEDV) Strain NJ Using Porcine Intestinal Epithelial Cells" Viruses 9, no. 1: 19. https://doi.org/10.3390/v9010019