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Low and high concentration lyophilized sera samples were used as quality controls.
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Sera and synovial fluid samples were used at a 1 100 dilution (a concentration selected after preliminary experiments).
Sera obtained from the blood samples were used for blood urea nitrogen (BUN) and serum creatinine determination using commercial kits (Ortho-Clinical Diagnostics, Rochester, NY, USA) according to the manufacturer's instructions.
A total of 226 duck sera samples were tested using the assay.
In contrast, no positive response was observed when the same sera samples were tested using peptide-based (peptides 5 and 11) ELISA.
IgG, IgG1 and IgG2a antibodies in sera samples were determined using an indirect ELISA assay according to the manufacture's instructions (R&D Systems, USA).
To demonstrate that the AuNP-based immunoassay can accurately quantify target molecules in biological samples, the patient sera samples were diluted using fetal bovine serum by 10 000-fold to correspond to the linear range of the calibration curve.
The molecular forms of TK1 in some of these sera samples were also investigated using size-exclusion chromatography, and important differences were revealed.
The collected sera samples were stored at −70 °C before being used in the study.
Sera samples were stored at −20 °C until used.
All sera samples were stored at -80°C until used.
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