Abstract
The frequent emergence of secondary infection and immunosuppression after porcine circovirus type 2 (PCV2) infection highlights the need to develop sensitive detection methods. A dual-signal amplification enzyme-linked immunosorbent assay (ELISA) based on a microplate coated with gold nanoparticle layers (GNPL) and tyramide signal amplification (TSA) was established. Results confirmed that the microplates coated with GNPL have a strong binding ability to the antibody without affecting the biological activity of the antibody. The microplates coated with GNPL have strong binding ability to the antibody, and the amplification of the tyramide signal is combined to further improve the sensitivity of PCV2. The PCV2 antibody does not crossreact with other viruses, demonstrating that the method has good specificity. A dual-signal amplification strategy is developed using microplates modified with GNPL and TSA to sensitively detect PCV2.
Highlights
Porcine circovirus type 2 (PCV2), which belongs to the genus Circovirus in the family Circoviridae, has been identified as the major etiological agent of postweaning multisystemic wasting syndrome (PMWS)
A dual amplification strategy was constructed by combining Tyramide signal amplification (TSA)- and gold nanoparticle layers (GNPL)-coated method could be used to sensitively quantify PCV2, and possibly detect low PCV2 levels from the microplates to develop a simple and sensitive enzyme-linked immunosorbent assay (ELISA) method for PCV2 detection
The unamplified, separate TSA signal amplification, separate GNPL signal amplification, and dual amplification (TSA + GNPL) methods were constructed separate GNPL signal amplification, and dual amplification (TSA + GNPL) methods were separately to verify the efficiency of dual signal amplification for PCV2 detection
Summary
Porcine circovirus type 2 (PCV2), which belongs to the genus Circovirus in the family Circoviridae, has been identified as the major etiological agent of postweaning multisystemic wasting syndrome (PMWS). A 100-fold sensitivity improvement was achieved by in situ hybridization by has been widely used in ELISA, flow cytometry, immunohistochemistry, and in situ hybridization usingfor biotin-labelled oligodeoxynucleotides and tyramide amplification in foot andand mouth the detection proteins, DNA, and pathogens because of signal its excellent amplification effects disease virus (FMDV) Adetection [15]. 100-fold sensitivity improvement wasaachieved by in situin hybridization by method following the tyramide signal amplification methodsignal that can detect porcine reproductive using biotin-labelled oligodeoxynucleotides and tyramide amplification in foot and mouth and diseasesyndrome virus (FMDV). A dual amplification strategy was constructed by combining TSA- and GNPL-coated method could be used to sensitively quantify PCV2, and possibly detect low PCV2 levels from the microplates to develop a simple and sensitive ELISA method for PCV2 detection. To the best of our knowledge, this study is the first report on the highly sensitive detection of PCV2
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