Complete Genome Sequence of K14JB01, a Novel Variant Strain of Porcine Epidemic Diarrhea Virus in South Korea

Cho, Yoon-Young; Lim, Suk-Kyung; Kim, Yong Kwan; Song, Jae‐Young; Lee, Joong‐Bok; An, Dong-Jun

doi:10.1128/genomea.00505-14

Cited by 18 publications

(15 citation statements)

References 8 publications

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“…Soluble expression of truncated S protein (tSc, aa 25-225) of PEDV CV777 prototype strain was reported in E. coli [20]. Here, we cloned and expressed NTD S25-229, an equivalent part of aa 25-225 of CV777 strain, of PEDV K14JB01 variant strain (Korean field strain isolated in 2013 [28]) in E. coli for our vaccine study following expression strategy of above report. The expression plasmid, pET28/S25-229, producing recombinant S25-229 protein (containing 205 amino acids, spanning amino acid region 25-229 of S protein) was constructed and transformed into the E. coli BL21 (DE3) strain.…”

Section: Confirmation Of Recombinant S25-229 Expression and Ibs Purifmentioning

confidence: 97%

Production of soluble truncated spike protein of porcine epidemic diarrhea virus from inclusion bodies of Escherichia coli through refolding

Piao

Lee

Bok

et al. 2016

Protein Expression and Purification

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The emergence of highly pathogenic variant porcine epidemic diarrhea virus (PEDV) strains, from 2013 to 2014, in North American and Asian countries have greatly threatened global swine industry. Therefore, development of effective vaccines against PEDV variant strains is urgently needed. Recently, it has been reported that the N-terminal domain (NTD) of S1 domain of PEDV spike protein is responsible for binding to the 5-N-acetylneuraminic acid (Neu5Ac), a possible sugar co-receptor. Therefore, the NTD of S1 domain could be an attractive target for the development of subunit vaccines. In this study, the NTD spanning amino acid residues 25-229 (S25-229) of S1 domain of PEDV variant strain was expressed in Escherichia coli BL21 (DE3) in the form of inclusion bodies (IBs). S25-229 IBs were solubilized in 20 mM sodium acetate (pH 4.5) buffer containing 8 M urea and 1 mM dithiothreitol with 95% yield. Solubilized S25-229 IBs were refolded by 10-fold flash dilution and purified by one-step cation exchange chromatography with >95% purity and 20% yield. The CD spectrum of S25-229 showed the characteristic pattern of alpha helical structure. In an indirect ELISA, purified S25-229 showed strong reactivity with mouse anti-PEDV sera. In addition, immunization of mice with 20 μg of purified S25-229 elicited highly potent serum IgG titers. Finally, mouse antisera against S25-229 showed immune reactivity with native PEDV S protein in an immunofluorescence assay. These results suggest that purified S25-229 may have potential to be used as a subunit vaccine against PEDV variant strains.

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Section: Confirmation Of Recombinant S25-229 Expression and Ibs Purifmentioning

confidence: 97%

Production of soluble truncated spike protein of porcine epidemic diarrhea virus from inclusion bodies of Escherichia coli through refolding

Piao

Lee

Bok

et al. 2016

Protein Expression and Purification

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“…The US PEDV strains identified during the initial outbreak in 2013 were closely related genetically to the Chinese strains (China/2012/ AH2012) reported in 2011-2012 Chen et al, 2014), indicating emergence of AH2012-like Chinese PEDV strains in the US. The US-like PEDV strains were also found in diarrheic piglets in South Korea and Taiwan during late 2013 and early 2014 (Cho et al, 2014;Lin et al, 2014), although whether Chinese or US PEDV strains could have been transmitted to pigs in South Korea and Taiwan is unknown. Further investigations are needed to clarify if Chinese or US PEDV strains were already present in South Korea and Taiwan before the related outbreaks were first identified.…”

Section: Molecular Epidemiology Of Pedv In the Us In 2013-2014mentioning

confidence: 99%

Porcine epidemic diarrhea virus infection: Etiology, epidemiology, pathogenesis and immunoprophylaxis

Jung

Saif

2015

The Veterinary Journal

406

418

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Porcine epidemic diarrhea virus (PEDV), a member of the genera Alphacoronavirus in the family Coronaviridae, causes acute diarrhea/vomiting, dehydration and high mortality in seronegative neonatal piglets. For the last three decades, PEDV infection has resulted in significant economic losses in the European and Asian pig industries, but in 2013-2014 the disease was also reported in the US, Canada and Mexico. The PED epidemic in the US, from April 2013 to the present, has led to the loss of more than 10% of the US pig population. The disappearance and re-emergence of epidemic PED indicates that the virus is able to escape from current vaccination protocols, biosecurity and control systems. Endemic PED is a significant problem, which is exacerbated by the emergence (or potential importation) of multiple PEDV variants. Epidemic PEDV strains spread rapidly and cause a high number of pig deaths. These strains are highly enteropathogenic and acutely infect villous epithelial cells of the entire small and large intestines although the jejunum and ileum are the primary sites. PEDV infections cause acute, severe atrophic enteritis accompanied by viremia that leads to profound diarrhea and vomiting, followed by extensive dehydration, which is the major cause of death in nursing piglets. A comprehensive understanding of the pathogenic characteristics of epidemic or endemic PEDV strains is needed to prevent and control the disease in affected regions and to develop an effective vaccine. This review focuses on the etiology, epidemiology, disease mechanisms and pathogenesis as well as immunoprophylaxis against PEDV infection.

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“…1). K14JB01 is a novel variant PEDV strain isolated from an infected pig in South Korea in 2013 (Cho et al, 2014). All of the reference panels were obtained from the Korea Veterinary Culture Collection (KVCC) (http://kvcc.kahis.go.kr) and were used to test the specificity of the developed IC assay.…”

Section: Reference Panel and Field Samplesmentioning

confidence: 99%

A novel diagnostic approach to detecting porcine epidemic diarrhea virus: The lateral immunochromatography assay

Kim

Lim

Cho

et al. 2015

Journal of Virological Methods

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Porcine epidemic diarrhea virus (PEDV) causes acute diarrhea and dehydration in sucking piglets and has a high mortality rate. An immunochromatography (IC) assay, known as a lateral flow test, is a simple device intended to detect the presence of target pathogens. Here, we developed an IC assay that detected PEDV antigens with 96.0% (218/227) sensitivity and 98.5% (262/266) specificity when compared with real-time reverse transcriptase (RT)-PCR using FAM-labeled probes based on sequences from nucleocapsid genes. The detection limits of the real-time RT-PCR and IC assays were 1×10(2) and 1×10(3) copies, respectively. The IC assay developed herein did not detect non-specific reactions with other viral or bacterial pathogens, and the assay could be stored at 4°C or room temperature for 15 months without affecting its efficacy. Thus, the IC assay may result in improved PED detection and control on farms, and is a viable alternative to current diagnostic tools for PEDV.

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Complete Genome Sequence of K14JB01, a Novel Variant Strain of Porcine Epidemic Diarrhea Virus in South Korea

Cited by 18 publications

References 8 publications

Production of soluble truncated spike protein of porcine epidemic diarrhea virus from inclusion bodies of Escherichia coli through refolding

Production of soluble truncated spike protein of porcine epidemic diarrhea virus from inclusion bodies of Escherichia coli through refolding

Porcine epidemic diarrhea virus infection: Etiology, epidemiology, pathogenesis and immunoprophylaxis

A novel diagnostic approach to detecting porcine epidemic diarrhea virus: The lateral immunochromatography assay

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