Gene encoding 22-380 aa of spike protein of porcine epidemic diarrhea virus (PEDV) isolate in China was cloned and expressed as a recombinant protein in Escherichia coli BL21(DE3). Female BALB/c mice were immunized with the purified recombinant S protein, and a monoclonal antibody (MAb) designated as 2D1 against S protein was achieved by hybridoma technique. MAb 2D1 reacted with S protein of PEDV specifically. The monoclonal antibody 2D1 may provide a useful tool as a specific diagnostic reagent for detecting S protein of the Chinese variant of PEDV and for further investigation into the virus' pathogenic mechanism.
Get full access to this article
View all access options for this article.
References
1.
SongDS, and ParkBK: Porcine epidemic diarrhea virus: a comprehensive review of molecular epidemiology, diagnosis, and vaccine. Virus Genes, 2012; 44:167–175.
2.
ParkSJ, SongDS, and ParkBK: Molecular epidemiology and phylogenetic analysis of porcine epidemic diarrhea virus (PEDV) field isolates in Korea. Arch Virol, 2013; 158:1533–1541.
3.
ChenJF, WangCB, ShiHY, QiuHJ, LiuSW, ChenXJ, ZhangZB, and FengL: Molecular epidemiology of porcine epidemic diarrhea virus in China. Arch Virol, 2010; 155:1471–1476.
4.
ChenX, YangJ, YuF, GeJ, LinT, and SongT: Molecular characterization and phylogenetic analysis of porcine epidemic diarrhea virus (PEDV) samples from field cases in Fujian, China. Virus Genes, 2012; 45:499–507.
5.
LiZ, ChenF, YuanY, ZengX, WeiZ, ZhuL, SunB, XieQ, CaoY, XueC, MaJ, and BeeY: Sequence and phylogenetic analysis of nucleocapsid genes of porcine epidemic diarrhea virus (PEDV) strains in China. Arch Virol, 2013; 158:1267–1273.
6.
LiWT, LiH, LiuYB, PanYF, DengF, SongYH, TangXB, and HeQG: New variants of porcine epidemic diarrhea virus, China. Emerg Infect Dis, 2011; 18:1350–1353.
7.
LiZL, ZhuL, MaJY, ZhouQF, SongYH, SunBL, ChenRA, XieQM, and BeeYZ: Molecular characterization and phylogenetic analysis of porcine epidemic diarrhea virus (PEDV) field strains in south China. Virus Genes, 2012; 45:181–185.
8.
FanJH, ZuoYZ, LiJH, and PeiLH: Heterogeneity in membrane protein genes of porcine epidemic diarrhea viruses isolated in China. Virus Genes, 2012; 45:113–117.
9.
SunRQ, CaiRJ, ChenYQ, LiangPS, ChenDK, and SongCX: Outbreak of porcine epidemic diarrhea in suckling piglets, China. Emerg Infect Dis, 2012; 18:161–163.
10.
StevensonGW, HoangH, SchwartzKJ, BurroughER, SunD, MadsonD, CooperVL, PillatzkiA, GaugerP, SchmittBJ, KosterLG, KillianML, and YoonKJ: Emergence of porcine epidemic diarrhea virus in the United States: clinical signs, lesions, and viral genomic sequences. J Vet Diag Invest, 2013; 25:649.
11.
ChenQ, LiG, StaskoJ, ThomasJT, StenslandWR, PillatzkiAE, GaugerPC, SchwartzKJ, MadsonD, YoonKJ, StevensonGW, BurroughER, HarmonKM, MainRG, and ZhangJ: Isolation and characterization of porcine epidemic diarrhea viruses associated with the 2013 disease outbreak in US Swine. J Clin Microbiol, 2014; 52:234–243.
12.
ShiratoK, MaejimaM, MatsuyamaS, UjikeM, MiyazakiA, TakeyamaN, IkedaH, and TaguchiF: Mutation in the cytoplasmic retrieval signal of porcine epidemic diarrhea virus spike (S) protein is responsible for enhanced fusion activity. Virus Res, 2011; 161:188–193.
13.
XiaoH, XuLH, YamadaY, and LiuDX: Coronavirus spike protein inhibits host cell translation by interaction with eIF3f. PLoS One, 2008; 3:e1494.
14.
LuisaDM, GabriellaM, LongoM, FilomenaF, SerenaM, ValentinaI, NicolaD, and UgoP: Bid cleavage, cytochrome c release and caspase activation in canine coronavirus-induced apoptosis. Vet Microbiol, 2010; 141:36–45.
15.
LiuaY, and ZhangXM: Murine coronavirus-induced oligodendrocyte apoptosis is mediated through the activation of the Fas signaling pathway. Virology, 2007; 360:364–375.
16.
DingL, XuXG, HuangY, LiZC, ZhangKZ, ChenGD, YuGS, WangZS, LiW, and TongDW: Transmissible gastroenteritis virus infection induces apoptosis through FasL- and mitochondria mediated pathways. Vet Microbiol, 2012; 158:12–22.
17.
ChowKY, YeungYS, HonCC, ZengF, LawKM, and LeungFC: Adenovirus-mediated expression of the C-terminal domain of SARS-CoV spike protein is sufficient to induce apoptosis in Vero E6 cells. FEBS Lett, 2005; 579(30):6699–6704.
18.
SunDB, FengL, and ShiHY: Spike protein region (aa 636-789) of porcine epidemic diarrhea virus is essential for induction of neutralizing antibodies. Acta Virol, 2007; 51:327–338.
19.
SunDB, FengL, ShiHY, ChenJF, CuiXC, ChenHY, LiuSW, TongYE, WangYF, and TongGZ: Identification of two novel B cell epitopes on porcine epidemic diarrhea virus spike protein. Vet Microbiol, 2008; 131:73–81.
20.
KangTJ, SeoJE, KimDH, KimTG, JangYS, and YangMS: Cloning and sequence analysis of the Korean strain of spike gene of porcine epidemic diarrhea virus and expression of its neutralizing epitope in plants. Prot Express Purif, 2005; 41:378–383.
21.
WangXM, NiuBB, YanH, GaoDS, YangX, ChenL, ChangHT, ZhaoJ, and WangCQ: Genetic properties of endemic Chinese porcine epidemic diarrhea virus strains isolated since 2010. Arch Virol, 2013; 158:2487–2494.
22.
WangG, WangY, LiCY, GaoTW, and LiuYF: Thiocyanate elution measurement of relative affinity of phage antibodies. Chin J Biotechnol, 2004; 20(3):429–433.
23.
CruzaDM, KimaCJ, and ShinHJ: The GPRLQPY motif located at the carboxyl-terminal of the spike protein induces antibodies that neutralize porcine epidemic diarrhea virus. Virus Res, 2008; 132:192–196.
24.
SatoTS, TakeyamaNS, KatsumataAS, TuchiyaKT, KodamaTS, and KusanagiKI: Mutations in the spike gene of porcine epidemic diarrhea virus associated with growth adaptation in vitro and attenuation of virulence in vivo. Virus, 2011; 43:72–78.
25.
KimSY, SongDS, and ParkBK: Differential detection of transmissible gastroenteritis virus and porcine epidemic diarrhea virus by duplex RT-PCR. J Vet Diagn Invest, 2001; 13:516–520.
26.
HanR, ShiXL, ZhaoPD, BaiJ, LiYF, WangXW, and JiangP: Development and application of an indirect ELISA method for detection of antibody to porcine epidemic diarrhea virus. Anim Husband Vet Med, 2013; 2(45):1–6.
27.
SozziE, LuppiA, LelliD, MartinAM, CanelliE, BrocchiE, LavazzaA, and CordioliP: Comparison of enzyme-linked immunosorbent assay and RT-PCR for the detection of porcine epidemic diarrhoea virus. Res Vet Sci, 2010; 31(1):166–168.
28.
ZhuWZ, WangXJ, and ChangJB: Detection of virus antigen in feces of PED-infected pigs with ELISA (double antibody sandwich method). Bull Vet Coll PLA, 1988; 2(8):127–130.
29.
SunDB, FengL, and ShiHY: Preparation and characterization of the monoclonal antibodies against neutralizing epitope region of PEDV S protein. Chin J Prevent Vet Med, 2007; 11(29):887–890.
