title: Lateral flow assay for rapid detection of white spot syndrome virus (WSSV) using a phage-displayed peptide as bio-recognition probe.
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.
authors.affiliation: Nanobioscience Group, Agharkar Research Institute, Pune, 411004, India.
authors.forename: Prabir Kumar
authors.initials: PK
authors.lastname: Kulabhusan
authors.affiliation: Nanobioscience Group, Agharkar Research Institute, Pune, 411004, India.
authors.forename: Jyutika M
authors.initials: JM
authors.lastname: Rajwade
authors.affiliation: OIE reference laboratory for WTD, C. Abdul Hakeem College, Melvisharam, Tamilnadu, 632509, India.
authors.forename: A S
authors.initials: AS
authors.lastname: Sahul Hameed
authors.affiliation: Nanobioscience Group, Agharkar Research Institute, Pune, 411004, India. kpaknikar@gmail.com.
authors.forename: Kishore M
authors.initials: KM
authors.lastname: Paknikar
mesh_terms: D000818:Animals; D017756:Aquaculture; D001435:Bacteriophages / Q000378:metabolism; D015153:Blotting, Western; D004279:DNA, Viral; D007194:India; D033561:Penaeidae / Q000821:virology*; D019151:Peptide Library; D010455:Peptides / Q000737:chemistry / Q000302:isolation & purification; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification; D014759:Viral Envelope Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*; D046848:White spot syndrome virus 1 / Q000737:chemistry / Q000302:isolation & purification*
chemical_list: D004279:DNA, Viral; D019151:Peptide Library; D010455:Peptides; D011994:Recombinant Proteins; C548473:VP28 protein, white spot syndrome virus; D014759:Viral Envelope Proteins
keywords: Lateral flow assay; Phage display peptides; White spot syndrome virus