An ELISA optimized for porcine epidemic diarrhoea virus detection in faeces

Rodák, L.; Valíček, L.; Smíd, B; Nevoránková, Z.

doi:10.1016/j.vetmic.2004.09.020

Cited by 37 publications

(43 citation statements)

References 24 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…On the other hand, PEDV N protein forms a helical nucleocapsid with genomic RNA and is the predominant antigen produced in coronavirus-infected cells, thus making it a major viral target. N protein was also applied as a sensitive diagnostic antigen to develop specific and sensitive ELISA (Rodak et al 2005;Hou et al 2007a).…”

Section: Discussionmentioning

confidence: 99%

Comparison of serum neutralization and enzyme-linked immunosorbent assay on sera from porcine epidemic diarrhea virus vaccinated pigs

Paudel

Park

Jang

et al. 2014

Veterinary Quarterly

View full text Add to dashboard Cite

Background: Porcine epidemic diarrhea virus (PEDV) is an economically important pathogen of swine. Objective: Serum neutralization (SN) and enzyme-linked immunosorbent assay (ELISA) test results as well as the utility of spike proteins S1, S2, and S3 and entire nucleocapsid protein were compared. Animals and methods: Serum samples from 400 pigs vaccinated against PEDV strain SM98P were collected from 78 farms in Korea. SN test and ELISA were performed to confirm the presence of antibodies. For prokaryotic expression of partial fragments of spike protein the size and location of S1, S2, and S3, and full nucleocapsid protein, polymerase chain reaction was performed using specific primers. Results:Comparison of these results demonstrated that there was a correlation between the SN and ELISA results. Sera with higher neutralizing activity also had higher IgG titer. The antibody profiling data presented the correlation of neutralizing activity with the level of spike protein antibody. In particular, the S3 region may have an important role in neutralizing activity. Conclusions: We confirmed that the carboxy-terminal region that includes the endodomain of the S protein induced stronger neutralizing activity than the region that includes the ectodomain. Clinical relevance: The region of the S protein may have a stronger neutralizing KPEDV-9 epitope and could be useful for the evaluation of future PEDV vaccine efficacy.

show abstract

Section: Discussionmentioning

confidence: 99%

Comparison of serum neutralization and enzyme-linked immunosorbent assay on sera from porcine epidemic diarrhea virus vaccinated pigs

Paudel

Park

Jang

et al. 2014

Veterinary Quarterly

View full text Add to dashboard Cite

show abstract

“…This was the case, in particular, for the diagnosis of viral gastroenteritis, for which molecular techniques capable of identifying most of the virus families involved in human gastroenteritis have been established . A similar shift in practice occurred in veterinary medicine, with ELISAs and PCR progressively replacing TEM for the routine diagnosis of viral infections . In human medicine, the use of TEM to differentiate between smallpox virus and the other viruses present in the fluids of cutaneous vesicles is no longer required, since the successful eradication of the variola virus in 1980 thanks to a worldwide vaccination program .…”

Section: Historymentioning

confidence: 99%

Virus detection by transmission electron microscopy: Still useful for diagnosis and a plus for biosafety

Roingeard

Raynal

Eymieux

et al. 2018

Reviews in Medical Virology

View full text Add to dashboard Cite

Summary Transmission electron microscopy (TEM) is the only imaging technique allowing the direct visualization of viruses, due to its nanometer‐scale resolution. Between the 1960s and 1990s, TEM contributed to the discovery of many types of viruses and served as a diagnostic tool for identifying viruses directly in biological samples, either in suspension or in sections of tissues or mammalian cells grown in vitro in contact with clinical samples. The diagnosis of viral infections improved considerably during the 1990s, with the advent of highly sensitive techniques, such as enzyme‐linked immunosorbent assay (ELISA) and PCR, rendering TEM obsolete for this purpose. However, the last 20 years have demonstrated the utility of this technique in particular situations, due to its “catch‐all” nature, making diagnosis possible through visualization of the virus, without the need of prior assumptions about the infectious agent sought. Thus, in several major outbreaks in which molecular techniques failed to identify the infectious agent, TEM provided the answer. TEM is also still occasionally used in routine diagnosis to characterize infections not diagnosed by molecular assays. It is also used to check the microbiological safety of biological products. Many biopharmaceuticals are produced in animal cells that might contain little‐known, difficult‐to‐detect viruses. In this context, the “catch‐all” properties of TEM make it possible to document the presence of viruses or virus‐like particles in these products.

show abstract

“…It has become one of the severe viral diarrhea diseases that lead to serious economic losses in swine-raising countries (Debouck et al, 1982;Hofmann and Wyler, 1987). Although some currently established methods for serodiagnosis of PED virus infection had been reported, most of all their detecting reagents are all prepared with virus antigen from PED virus infected Vero cells or intestine and fecal samples of experimentally infected sows (Jin et al, 2004(Jin et al, , 2005Rodak et al, 2005). However, the narrow infection spectrum and the lower cell infection titre of PED virus limited the yield in laboratory, and the clinical application of PED whole virus antigen presents potential risk.…”

Section: Introductionmentioning

confidence: 99%

High-level prokaryotic expression of envelope exterior of membrane protein of porcine epidemic diarrhea virus

Gao

Zha

Baoxian

et al. 2007

Veterinary Microbiology

View full text Add to dashboard Cite

The truncated fragment M' gene, encoding the exterior of the viral envelope protein of PEDV, was subcloned into prokaryotic expression vector pGEX-6p-1. The recombinant plasmid pGEX-6p-M' was constructed and transformed into E. coli BL21(DE3)pLysS for expression. SDS-PAGE analysis showed recombinant truncated M' protein was highly expressed by pGEX-6p-M' and the product fusion protein GST-M' reached 45% in the total bacteria proteins with the analysis of software AlphaImager2200. The preliminary purified recombinant protein was evaluated for its antigenicity and reactivity through Western blotting and indirect enzyme-linked immunosorbent assay (ELISA) with monoclonal antibody against M protein of PEDV and porcine polyclonal anti-PEDV antiserum as the primary antibody. The results indicated the recombinant truncated M' protein should be candidate as a feasible recombinant diagnostic reagent.

show abstract

An ELISA optimized for porcine epidemic diarrhoea virus detection in faeces

Cited by 37 publications

References 24 publications

Comparison of serum neutralization and enzyme-linked immunosorbent assay on sera from porcine epidemic diarrhea virus vaccinated pigs

Comparison of serum neutralization and enzyme-linked immunosorbent assay on sera from porcine epidemic diarrhea virus vaccinated pigs

Virus detection by transmission electron microscopy: Still useful for diagnosis and a plus for biosafety

High-level prokaryotic expression of envelope exterior of membrane protein of porcine epidemic diarrhea virus

Contact Info

Product

Resources

About