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

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• 作者：Niels Gr&uuml ; tzner^a ; ¹ ; ^{tierarztbz@hotmail.com" class="auth_mail} ; Romy M. Heilmann^a ; ¹ ; Jan S. Suchodolski^a ; Jö ; rg M. Steiner^a ; Andreas Holzenburg^b ; ^c ; ^d
• 关键词：Calprotectin ; Cold-microwave technology ; ELISA ; Feces ; Serum ; Validation
• 刊名：Analytical Biochemistry
• 出版年：15 July 2014
• 年：2014
• 卷：457
• 期：Complete
• 页码：65-73
• 全文大小：1756 K

文摘

The enzyme-linked immunosorbent assay (ELISA) constitutes an important clinical diagnostic approach. However, the prolonged incubation times involved lead to turnaround times of typically 猢? 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 猢?7.2%. For samples run on precoated ELISA plates over 33 days %CVs were 猢?2.5%. 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.