Nanoparticle-based lateral flow biosensor for visual detection of fish nervous necrosis virus amplification products

Toubanaki, Dimitra K.; Margaroni, Maritsa; Karagouni, Evdokia

doi:10.1016/j.mcp.2015.03.005

Cited by 29 publications

(12 citation statements)

References 53 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Figure 2b proves that all PBS stabilized particles were successfully conjugated with antibodies while no conjugation occurred on the citrate 40 nm nanoparticles. Therefore, Au NPs stabilized with PBS buffer facilitates proteins (antibodies) conjugation while the citrate buffer is more efficient on conjugation of SH-tailed oligonucleotides 8 . Use of 5 and 60 nm NPs resulted in faint control zones.…”

Section: Resultsmentioning

confidence: 99%

“…The visual detection was completed within 30 min. Longer times did not affect the assay results 8 . After completion of the assay, the LFBs were scanned with a desktop scanner and the band densities were quantified with ImageJ software 30 .…”

Section: Lateral Flow Biosensor Detection Assay Of Nodavirus Virionsmentioning

confidence: 90%

“…The membrane was dried in an oven for 1 h at 80 °C, and the sensors were assembled as described before. LFBs with a 4-mm width were cut using a Guillotine cutter and stored dry, at ambient temperature 8,26 .…”

Section: Preparation Of Anti-nodavirus Antibody Conjugated Aumentioning

confidence: 99%

See 2 more Smart Citations

Development of a Nanoparticle-based Lateral Flow Strip Biosensor for Visual Detection of Whole Nervous Necrosis Virus Particles

Toubanaki

Margaroni

Prapas³

et al. 2020

Sci Rep

Self Cite

View full text Add to dashboard Cite

Effective analysis of pathogens causing human and veterinary diseases demands rapid, specific and sensitive detection methods which can be applied in research laboratory setups and in field for routine diagnosis. Paper lateral flow biosensors (LFBs) have been established as attractive tools for such analytical applications. In the present study a prototype LFB was designed for whole particles (virions) detection of nodavirus or fish nervous necrosis virus. Nodavirus is an important threat in the aquaculture industry, causing severe economic losses and environmental problems. The LFB was based on polyclonal antibodies conjugated on gold nanoparticles for signal visualization. Brain and retinas from fish samples were homogenized, centrifuged and the supernatant was directly applied on the LFB. formation of a red test line was indicative of nodavirus virions presence. nodavirus visual detection was completed in short time (30 min). Key factors of the LFB development influencing the assays' detection limit were characterized and the optimum parameters were determined, enabling increased efficiency, excluding non-specific interactions. Therefore, the proposed LFB assay consists a robust, simple, low cost and accurate method for detection of nodavirus virions in fish samples. The proposed biosensor is ideal for development of a commercial kit to be used on aquaculture facilities by fish farmers. It is anticipated that disease monitoring and environmental safety will benefit from the simplification of time consuming and costly procedures.Aquaculture is essential to cover fish-product demands, providing seafood in high quantities, and covering more than the half amount of fish consumed worldwide. This fact drives a strong demand for high production efficiency in aquaculture industry in order to cover the feeding needs of the world's growing population, in the middle of an increasing environmental crisis 1,2 . As a result, the aquaculture industry has continuously increased profits in a high rate. However, outbreaks of diseases caused by infectious agents are significantly restricting intensified aquaculture. According to literature 3 , 22.6% of all disease outbreaks are caused by viruses. Among these, viral nervous necrosis (VNN), also named vacuolating encephalopathy and retinopathy or encephalomyelitis, is a devastating disease, which induces cell necrosis accompanied by vacuolation in fish retina and brain. Its clinical symptoms include changes in skin color with abnormal swimming, low feed ingestion and altered buoyancy in affected fish. The disease is caused by nervous necrosis virus (NNV) or nodavirus, affecting more than 30 different fish species, worldwide. VNN causes high mortalities (80-100% in several species e.g. European sea bass), emerging as a major problem especially in the Mediterranean area, since it cannot be prevented by vaccination or effective treatment 4-6 .Fish nervous necrosis virus (belonging to Betanodavirus genus and Nodaviridae family) is icosahedral, and non-enveloped (∼25 nm in diameter). I...

show abstract

Section: Resultsmentioning

confidence: 99%

Section: Lateral Flow Biosensor Detection Assay Of Nodavirus Virionsmentioning

confidence: 90%

See 1 more Smart Citation

Development of a Nanoparticle-based Lateral Flow Strip Biosensor for Visual Detection of Whole Nervous Necrosis Virus Particles

Toubanaki

Margaroni

Prapas³

et al. 2020

Sci Rep

Self Cite

View full text Add to dashboard Cite

show abstract

“…Further developments of the PCR technique have led to a PCR method that allows RGNNV and SJNNV genotyping [273] and to a nanoparticle-based paper lateral flow biosensor (LFB) for visual detection of RT-PCR products in biological samples using gold nanoparticles with a detection limit of 270 pg [281]. Lateral flow paper biosensors have demonstrated to be attractive analytical platform for detection of pathogens because they allow and accurate, rapid and sensitive diagnostics and are also appropriate for field analysis [281].…”

Section: Diagnostic Proceduresmentioning

confidence: 99%

“…Different nanostructures are available at present, such as nanoparticles (NP), carbon nanotubes (CNT), dendrimers, and quantum dots (QDs) among others, and can be applied to the identification of nucleic acids, proteins and viral particles as well as antibodies [307,308]. To the best of our knowledge, nano-based diagnostic methods have only been used twice for the detection of fish viruses-the infectious pancreatic necrosis virus [309] and NNV [281]. The use of a nanoparticle-based LFB for the detection of NNV amplification products enables accurate and fast diagnostics under field conditions, although does not increase sensitivity compared to RT-qPCR assays.…”

Section: Diagnostic Proceduresmentioning

confidence: 99%

Betanodavirus and VER Disease: A 30-year Research Review

2020

View full text Add to dashboard Cite

The outbreaks of viral encephalopathy and retinopathy (VER), caused by nervous necrosis virus (NNV), represent one of the main infectious threats for marine aquaculture worldwide. Since the first description of the disease at the end of the 1980s, a considerable amount of research has gone into understanding the mechanisms involved in fish infection, developing reliable diagnostic methods, and control measures, and several comprehensive reviews have been published to date. This review focuses on host-virus interaction and epidemiological aspects, comprising viral distribution and transmission as well as the continuously increasing host range (177 susceptible marine species and epizootic outbreaks reported in 62 of them), with special emphasis on genotypes and the effect of global warming on NNV infection, but also including the latest findings in the NNV life cycle and virulence as well as diagnostic methods and VER disease control.

show abstract