Selection and characterization of ssDNA aptamer targeting Macrobrachium rosenbergii nodavirus capsid protein: A potential capture agent in gold-nanoparticle-based aptasensor for viral protein detection

J Fish Dis. 2024 Feb;47(2):e13892. doi: 10.1111/jfd.13892. Epub 2023 Nov 28.

Abstract

The giant freshwater prawn holds a significant position as a valuable crustacean species cultivated in the aquaculture industry, particularly well-known and demanded among the Southeast Asian countries. Aquaculture production of this species has been impacted by Macrobrachium rosenbergii nodavirus (MrNV) infection, which particularly affects the larvae and post-larvae stages of the prawn. The infection has been recorded to cause mortality rates of up to 100% among the affected prawns. A simple, fast, and easy to deploy on-site detection or diagnostic method is crucial for early detection of MrNV to control the disease outbreak. In the present study, novel single-stranded DNA aptamers targeting the MrNV capsid protein were identified using the systematic evolution of ligands by exponential enrichment (SELEX) approach. The aptamer was then conjugated with the citrate-capped gold nanoparticles (AuNPs), and the sensitivity of this AuNP-based aptasensor for the detection of MrNV capsid protein was evaluated. Findings revealed that the aptamer candidate, APT-MrNV-CP-1 was enriched throughout the SELEX cycle 4, 9, and 12 with the sequence percentage of 1.76%, 9.09%, and 12.42%, respectively. The conjugation of APT-MrNV-CP-1 with citrate-capped AuNPs exhibited the highest sensitivity in detecting the MrNV capsid protein, where the presence of 62.5 nM of the viral capsid protein led to a significant agglomeration of the AuNPs. This study demonstrated the practicality of an AuNP-based aptasensor for disease diagnosis, particularly for detecting MrNV infection in giant freshwater prawns.

Keywords: Macrobrachium rosenbergii nodavirus; gold nanoparticle; ssDNA aptamer; viral capsid protein.

MeSH terms

  • Animals
  • Capsid Proteins / genetics
  • Capsid Proteins / metabolism
  • Citrates / metabolism
  • DNA, Single-Stranded
  • Fish Diseases* / diagnosis
  • Gold
  • Metal Nanoparticles*
  • Nodaviridae* / genetics
  • Palaemonidae* / genetics
  • Viral Proteins / genetics

Substances

  • Capsid Proteins
  • Viral Proteins
  • Gold
  • DNA, Single-Stranded
  • Citrates