Cold-microwave enhanced enzyme-linked immunosorbent assays--a path to high-throughput clinical diagnostics.
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.
Gastrointestinal Laboratory, Department of Small Animal Clinical Sciences, Texas A&M University, College Station, TX 77843, USA. Electronic address: tierarztbz@hotmail.com.
Niels
N
Grützner
Gastrointestinal Laboratory, Department of Small Animal Clinical Sciences, Texas A&M University, College Station, TX 77843, USA.
Romy M
RM
Heilmann
Gastrointestinal Laboratory, Department of Small Animal Clinical Sciences, Texas A&M University, College Station, TX 77843, USA.
Jan S
JS
Suchodolski
Gastrointestinal Laboratory, Department of Small Animal Clinical Sciences, Texas A&M University, College Station, TX 77843, USA.
Jörg M
JM
Steiner
Microscopy and Imaging Center, Texas A&M University, College Station, TX 77843, USA; Department of Biology, Texas A&M University, College Station, TX 77843, USA; Department of Biochemistry and Biophysics, Texas A&M University, College Station, TX 77843, USA.
Andreas
A
Holzenburg
D000818:Animals; D003080:Cold Temperature; D004285:Dogs; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005243:Feces / Q000737:chemistry; D006801:Humans; D039841:Leukocyte L1 Antigen Complex / Q000097:blood* / Q000737:chemistry*; D008872:Microwaves; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D039841:Leukocyte L1 Antigen Complex
Calprotectin; Cold-microwave technology; ELISA; Feces; Serum; Validation