Tangential flow ultrafiltration for detection of white spot syndrome virus (WSSV) in shrimp pond water

Alavandi, S.V.; Bharathi, R. Ananda; Kumar, Sujeet; Dineshkumar, N.; Saravanakumar, C.; Rajan, J.J.S.

doi:10.1016/j.jviromet.2015.03.001

Cited by 9 publications

(6 citation statements)

References 33 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…The interesting point here is that we could floc the virus by alum in less volume of infected water, which is much lesser compared to the report of >60 L water for WSSV concentration and detection by PCR [21]. In addition to alum, several other agents have been tried by others to concentrate the virus, including TFF with skimmed-milk for adenovirus [14], glass-fiber filter pre-coated with bovine serum albumin for marine birnavirus [15], TFF for infectious salmon anemia virus [16] and TFF for WSSV [21]. However, these methods require laboratory, specific skills and equipment, which further makes the detection method costlier, time consuming and complex.…”

Section: Discussionmentioning

confidence: 84%

“…Centrifugation and membrane filtration, capacitive biosensor, combined ferric colloid adsorption and foam separation-based methods were employed to concentrate the WSSV in sea water for the detection by PCR [17-20]. In the recent study, TFF was used to concentrate the WSSV in shrimp pond water followed by PCR detection [21]. However, these methods are complex, sophisticated and costly for the field level detection of WSSV in shrimp culture.…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Concentrating white spot syndrome virus by alum for field detection using a monoclonal antibody based flow-through assay

et al. 2016

View full text Add to dashboard Cite

A simple and easy method of concentrating white spot syndrome virus by employing aluminium sulphate, alum as a flocculant was developed and evaluated for field detection. The concentrated virus was detected by a monoclonal antibody based flow-through assay, RapiDot and compared its performance with polymerase chain reaction. The semi-purified virus that was flocculated by 15 and 30 ppm alum in a 50 ml cylinder can be detected successfully by both RapiDot and I step PCR. In addition, alum could also flocculate the virus that is detectable by II step PCR and the concentration of virus was similar to the one observed in water from an infected pond. Furthermore, experimental infection studies validated the successful concentration of virus by alum flocculation followed by rapid detection of virus using monoclonal antibody based RapiDot. Overall, the results obtained in this study indicate that the white spot syndrome virus in water could be easily flocculated by alum for sensitive field detection by monoclonal antibody based RapiDot.

show abstract

Section: Discussionmentioning

confidence: 84%

Section: Introductionmentioning

confidence: 99%

Concentrating white spot syndrome virus by alum for field detection using a monoclonal antibody based flow-through assay

et al. 2016

View full text Add to dashboard Cite

show abstract

“…groundwater and tap water need about 100 L while recreational water 10 L for processing. In another study, WSSV was concentrated and detected by PCR after >60 L of water was processed [24]. However, this study has demonstrated that WSSV can be concentrated by skim milk flocculation from less volume of infected water for detection.…”

Section: Discussionmentioning

confidence: 85%

“…Virus concentration and detection methods in water are many [18,22,23] however protocols that have been developed for WSSV include tangential flow ultra filtration [24] followed by nested PCR (Polymerase Chain Reaction) and qPCR detection, combined ferric colloid adsorption and foam separation [25] followed by PCR detection, centrifugation [26] and membrane filtration [27] followed by PCR detection. Recently, [28] used inorganic flocculent, alum at 15 and 30 ppm to concentrate WSSV from water and detected successfully by monoclonal antibody based flow-through assay, RapiDot and PCR.…”

Section: Introductionmentioning

confidence: 99%

Skim Milk Flocculation Concentrates White Spot Syndrome Virus in Seawater for Detection Using A Monoclonal Antibody Based Flow-Through Assay

Ghambi¹,

Kalkuli²,

Patil³

et al. 2018

IJPFS

View full text Add to dashboard Cite

Since its emergence, white spot syndrome virus has caused huge loss to shrimp aquaculture. However, these viruses are present in very low copy number in water and hence, concentrating them prior to detection is very much essential. In the present study, skim milk (0.01%) was employed for one step concentration of white spot syndrome virus from seawater under acidic (pH 3.5) and neutral (pH 7.4) conditions. The concentrated virus was detected using a monoclonal antibody based flow-through assay, RapiDot and compared its performance with Polymerase Chain Reaction (PCR). In an invitro experiment, the recombinant VP28 protein of white spot syndrome virus was flocculated successfully by skim milk in 50 mL measuring cylinders and detectable in both sub-surface and floc samples by RapiDot. Further, these results were validated by experimental infection studies using virus infected water for skim milk flocculation and detected the virus in floc samples 6 h earlier by RapiDot, compared to 1-step PCR. Overall, a simple method to concentrate white spot syndrome virus in water by skim milk flocculation for detection by RapiDot has been developed and may be adaptable to field conditions.

show abstract

“…Los anticuerpos no unidos fueron removidos mediante lavado tres veces con Buffer sodio-fosfato [11]. procesar 100 L de agua, los investigadores obtuvieron un volumen final de 100 mL y un rango de recuperación viral que osciló entre un 7.5 a 89.61% [14]. En la figura 2 se aprecia con claridad que el contenido de RNA viral recuperado empleando UFT es menor al obtenido empleando IMS.…”

Section: Funcionalización De Las Partículas Magnéticasunclassified

Comparación De Dos Técnicas De Concentración Viral a Partir De Aguas Crudas Del Departamento Norte De Santander

Villamizar

Ortiz

2017

BISTUA

View full text Add to dashboard Cite

ResumenLos métodos de detección de virus entéricos a nivel de laboratorio presentan cierto nivel de complejidad, debido a que estos entes se encuentran muy diluidos en aguas. Esto genera la necesidad de encontrar métodos eficientes que permitan concentrarlos de forma rápida, selectiva, sensible, eliminando interferencias presentes en la matriz y con la certeza de tener partículas virales infecciosas. En este estudio se comparó la ultrafiltración tangencial (UFT) y la separación inmunomagnética (IMS) como técnicas de concentración viral, empleando Rotavirus como modelo. El grado de eficiencia de las mismas fue confirmado empleando la reacción en cadena de la polimerasa acoplada a una reversotranscriptasa (RT-PCR) la cual facilitó la identificación de Rotavirus a partir de aguas crudas en cuatro plantas de potabilización del departamento Norte de Santander, incluyendo sus fuentes de abastecimiento. La UFT se llevó a cabo en un equipo dotado entre otros componentes de membranas millipore de 0.8, 0.45 and 0.22 µm. Para la IMS se emplearon micropartículas de 2.0 µm funcionalizadas con anticuerpos monoclonales anti-Rotavirus.Los resultados permitieron determinar que tanto la IMS como la UFT concentraron eficientemente Rotavirus a partir de agua crudas. No obstante, la IMS exhibió mayor sensibilidad a la hora de comparar la cantidad de RNA viral extraído a partir de cada muestra analizada.Además, está técnica sólo requirió 2 horas para obtener el concentrado viral en comparación de 24 horas empleadas por la UFT. Por tanto, se recomienda emplear para la concentración de virus patógenos presentes en aguas crudas la técnica de separación inmunomagnética. Usualmente, los virus se encuentran muy diluidos en agua (1 a 10 3 partículas virales/Lt de agua) [2,5,6], creando la necesidad de pretratamientos de la muestra para poder concentrar las partículas virales. Existen diferentes técnicas de concentración entre las cuales se encuentra la ultrafiltración tangencial (UFT).Esta técnica consiste en realizar una serie de filtrados en membranas millipore de distinto tamaño, dirigiendo la muestra a un filtro de flujo tangencial por medio de una bomba peristáltica de alta presión, produciendo dos fracciones, la filtrada de interés y la excluida por el filtro [7]. Posteriormente, se realiza una segunda ultrafiltración, de donde se obtiene el filtrado final que es analizado a través de técnicas de biología molecular para realizar la respectiva detección del patógeno [8].No obstante, esta técnica requiere el uso de métodos adicionales a los moleculares para poder determinar el grado de infectividad de las partículas virales concentradas.Otra técnica de concentración viral es la separación inmunomagnética (IMS), la cual emplea partículas magnéticas acopladas a anticuerpos capaces de detectar, concentrar y separar patógenos. La afinidad existente entre el antígeno y el anticuerpo, los cuales se unen a través de interacciones supramoleculares mediado principalmente por puentes de hidrógenos, uniones iónicas y fuerzas de van der Waals [9] h...

show abstract

Tangential flow ultrafiltration for detection of white spot syndrome virus (WSSV) in shrimp pond water

Cited by 9 publications

References 33 publications

Concentrating white spot syndrome virus by alum for field detection using a monoclonal antibody based flow-through assay

Concentrating white spot syndrome virus by alum for field detection using a monoclonal antibody based flow-through assay

Skim Milk Flocculation Concentrates White Spot Syndrome Virus in Seawater for Detection Using A Monoclonal Antibody Based Flow-Through Assay

Comparación De Dos Técnicas De Concentración Viral a Partir De Aguas Crudas Del Departamento Norte De Santander

Contact Info

Product

Resources

About