Abstract
A sensitive and label-free analytical approach for the detection of porcine circovirus type 2 (PCV2) instead of PCV2 antibody in serum sample was systematically investigated in this research based on surface plasmon resonance (SPR) with an establishment of special molecular identification membrane. The experimental device for constructing the biosensing analyzer is composed of an integrated biosensor, a home-made microfluidic module, and an electrical control circuit incorporated with a photoelectric converter. In order to detect the PCV2 using the surface plasmon resonance immunoassay, the mercaptopropionic acid has been used to bind the Au film in advance through the known form of the strong S-Au covalent bonds formed by the chemical radical of the mercaptopropionic acid and the Au film. PCV2 antibodies were bonded with the mercaptopropionic acid by covalent -CO-NH- amide bonding. For the purpose of evaluating the performance of this approach, the known concentrations of PCV2 Cap protein of 10 µg/mL, 7.5 µg/mL, 5 µg/mL, 2.5 µg/mL, 1 µg/mL, and 0.5 µg/mL were prepared by diluting with PBS successively and then the delta response units (ΔRUs) were measured individually. Using the data collected from the linear CCD array, the ΔRUs gave a linear response over a wide concentration range of standard known concentrations of PCV2 Cap protein with the R-Squared value of 0.99625. The theoretical limit of detection was calculated to be 0.04 µg/mL for the surface plasmon resonance biosensing approach. Correspondingly, the recovery rate ranged from 81.0% to 89.3% was obtained. In contrast to the PCV2 detection kits, this surface plasmon resonance biosensing system was validated through linearity, precision and recovery, which demonstrated that the surface plasmon resonance immunoassay is reliable and robust. It was concluded that the detection method which is associated with biomembrane properties is expected to contribute much to determine the PCV2 in sample solutions instead of PCV2 antibody in serum samples quantitatively.
Highlights
Porcine circovirus (PCV) can be categorized into two genotypes: porcine circovirus type 1 and porcine circovirus type 2 according to the pathogenicity, antigenicity and nucleotide sequence
Porcine circovirus type 2 (PCV2) infection has been reported in many countries with a significant commercial production industry, which leads to high production losses for producers [1,2]
This paper presented a non-destructive, label free, and real-time optical surface plasmon resonance (SPR) immunoassay for detecting the porcine circovirus type 2 (PCV2) with the biological membrane adopted predominantly a self-assembly, which provides a breakthrough method for detecting viruses in PCV2-contained sample solutions, such as the PCV2 separated from the animal lymph nodes, manure or blenna narium
Summary
Porcine circovirus (PCV) can be categorized into two genotypes: porcine circovirus type 1 (no pathogenicity) and porcine circovirus type 2 (pathogenicity) according to the pathogenicity, antigenicity and nucleotide sequence. Porcine circovirus type 2 (PCV2) infection has been reported in many countries with a significant commercial production industry, which leads to high production losses for producers [1,2]. This PCV2 was closely related to post weaning multisystemic wasting syndrome (PMWS) disease which generally occurred worldwide [3,4]. A fast, sensitive and indirect ELISA method for the detection of the specificity, repeatability and stability of the PCV2 antibody of pigs has been established systematically. The colloidal gold method was used to detect the PCV2, including preparing gold labeled beads by spraying colloidal gold on the glass fiber for staphylococcal protein A (SpA), a type I membrane protein from the bacterium staphylococcus aureus, and producing colloidal gold immunochromatographic test strip for the detection of PCV2 antibodies has been constructed [12,13]
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