Abstract
The aim of the current study is to introduce a methodology aimed at producing a biosensor that uses gold nanoparticles (AuNPs) to detect porcine circovirus 2 (PCV-2). This biosensor was based on AuNPs, which were modified with self-assembled monolayers (SAMs) and antibodies. The AuNPs’ surface and virus modification process applied to enable antibody binding was accompanied by localized surface plasmon resonance (LSPR), surface plasmon resonance (SPR), transmission electron microscopy (TEM), and energy-dispersive X-ray spectroscopy (EDX). Virus quantification was possible by the light absorption difference in the spectrum at concentrations of 105, 106, 107, 108, and 109 DNA copies/mL PCV-2 in relation to quantitative PCR (qPCR), with an R2 value >0.98. The visualization of colorimetric changes in the different PCV-2 concentrations was possible without the use of equipment. The biosensor production methodology presented reproducibility and specificity, as well as easy synthesis and low cost. An enhanced version of it may be used in the future to replace traditional tests such as PCR.
Highlights
Porcine circovirus diseases (PCVD) is one of the most important diseases affecting domestic swine production; it is caused by porcine circovirus 2 (PCV-2) belonging to the family Circoviridae, genus
PCV-2b was the most prevalent after PCV-2a, and today, PCV-2d is widespread in many countries, suggesting a change of genotype from PCV-2b to PCV-2d, which is responsible for most clinical cases of postweaning multisystemic wasting syndrome (PMWS) [5,6]
All of the a biosensor that uses gold nanoparticles (AuNPs) surface modification stage was monitored through the localized surface plasmon resonance (LSPR) spectroscopy technique; they presented absorbance peak reduction and wavelength shift (Figure 1)
Summary
Porcine circovirus diseases (PCVD) is one of the most important diseases affecting domestic swine production; it is caused by porcine circovirus 2 (PCV-2) belonging to the family Circoviridae, genusCircovirus, which comprises three endemic types, PCV-1, PCV-2, and PCV-3 [1]. PCV-2 is the most relevant pathogen affecting pigs; it accounts for major damages to the economy, since the high mortality rate resulting from it causes irreparable damage losses to farmers [2]. The PCV-2 has a single-stranded DNA genome, with 1760 bases, wrapped by an icosaedric capsid [3]. This virus, which is approximately 17 nm diameter, is the agent responsible for several pig syndromes [4]. PCV-2b was the most prevalent after PCV-2a, and today, PCV-2d is widespread in many countries, suggesting a change of genotype from PCV-2b to PCV-2d, which is responsible for most clinical cases of postweaning multisystemic wasting syndrome (PMWS) [5,6]. The emergence of different genotypes derives from nucleotide additions or substitutions
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