Cold-microwave enhanced enzyme-linked immunosorbent assays—A path to high-throughput clinical diagnostics

Autor: Andreas Holzenburg, Romy M. Heilmann, Jan S. Suchodolski, Jörg M. Steiner, Niels Grützner
Rok vydání: 2014
Předmět:
Zdroj: Analytical Biochemistry. 457:65-73
ISSN: 0003-2697
Popis: The enzyme-linked immunosorbent assay (ELISA) constitutes an important clinical diagnostic approach. However, the prolonged incubation times involved lead to turnaround times of typically ⩾1 day, potentially delaying a definitive diagnosis or an adequate treatment plan for individual patients. Here cold-microwave technology (CMT) was employed to significantly reduce the times required for diagnostic ELISAs. The new approach was validated and compared to a conventional ELISA setup measuring canine calprotectin (cCP). Canine serum and fecal specimens were used for the analytical validation of cCP ELISA by conventional and CMT–ELISA. Cross-validation of both ELISA methods consisted of the determination of analytic sensitivity, linearity, accuracy, precision, and reproducibility. The long-term stability of antibody-coated ELISA plates was also evaluated up to 33 days. The ELISA approaches were comparable to each other. The observed-to-expected ratios for linearity and accuracy were 100.2 ± 11.8 and 98.1 ± 10.8% (mean ± standard deviation), respectively. Precision and reproducibility were ⩽17.2%. For samples run on precoated ELISA plates over 33 days %CVs were ⩽12.5%. While both ELISA approaches were analytically sensitive, linear, accurate, precise, and reproducible with measurements of cCP concentrations, CMT–ELISA offered a reduction in incubation times by 90–95%, facilitating a very fast turnaround time and suggesting CMT–ELISA for improved human and veterinary clinical diagnostics.
Databáze: OpenAIRE
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