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Lateral flow assay for rapid detection of white spot syndrome virus (WSSV) using a phage-displayed peptide as bio-recognition probe

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Abstract

White spot disease caused by the white spot syndrome virus (WSSV) has a major socio-economic impact on shrimp farming in India. It has been realized that a field-usable diagnostic capable of rapid detection of WSSV can prevent huge economic losses in disease outbreaks. In this work, we explored the possibility of using a peptide as bio-recognition probe in a field-usable device for the detection of WSSV from infected shrimps and prawns. A commercially available random phage-display library was screened against rVP28 (a major structural protein of WSSV, expressed as a recombinant protein in Escherichia coli). A bacteriophage clone VP28-4L was obtained, and its binding to purified rVP28 protein as well as WSSV from infected shrimp Litopaeneus vannamei tissue was confirmed by ELISA and western blot. The apparent equilibrium dissociation constant (Kd,app) was calculated to be 810 nM. VP28-4L did not show cross-reactivity with any other shrimp viruses. A 12-mer peptide (pep28, with the sequence ′TFQAFDLSPFPS′) displayed on the VP28-4L was synthesized, and its diagnostic potential was evaluated in a lateral flow assay (LFA). Visual detection of WSSV could be achieved using biotinylated-pep28 and streptavidin-conjugated gold nanoparticles. In LFA, 12.5 μg/mL of the virus could be detected from L. vannamei gill tissue homogenate within 20 min. Pep28 thus becomes an attractive candidate in bio-recognition of WSSV in field-usable diagnostic platforms benefitting the aquaculture sector.

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Acknowledgments

PK is thankful to the Indian Council of Medical Research (ICMR), Government of India for awarding a research fellowship.

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Authors and Affiliations

  1. Nanobioscience Group, Agharkar Research Institute, Pune, 411004, India

    Prabir Kumar Kulabhusan, Jyutika M. Rajwade & Kishore M. Paknikar

  2. OIE reference laboratory for WTD, C. Abdul Hakeem College, Melvisharam, Tamilnadu, 632509, India

    A. S. Sahul Hameed

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  1. Prabir Kumar Kulabhusan
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  2. Jyutika M. Rajwade
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  3. A. S. Sahul Hameed
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  4. Kishore M. Paknikar
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Correspondence to Kishore M. Paknikar.

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Ethics statement

Samples were collected as part of the National Surveillance Program for Aquatic Animal Diseases (NSPAAD) project; sub-project No. 24. The study was approved by the National Fisheries Development Board (NFDB)-Indian Council of Agricultural Research (ICAR), Government of India. The collection of samples was done as part of NSPAAD project; sub-project No. 24. Therefore, specific permissions were not required. The field studies involve only species cultivated in aquaculture farms/hatcheries.

Conflict of interest

PK declares that he has no conflict of interest.JMR declares that she has no conflict of interest. ASS declares that he has no conflict of interest. KMP declares that he has no conflict of interest. The authors declare that they have no conflict of interest.

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Kulabhusan, P.K., Rajwade, J.M., Sahul Hameed, A.S. et al. Lateral flow assay for rapid detection of white spot syndrome virus (WSSV) using a phage-displayed peptide as bio-recognition probe. Appl Microbiol Biotechnol 101, 4459–4469 (2017). https://doi.org/10.1007/s00253-017-8232-6

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  • DOI: https://doi.org/10.1007/s00253-017-8232-6

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