Format

Send to

Choose Destination
Anal Biochem. 2014 Jul 15;457:65-73. doi: 10.1016/j.ab.2014.04.026. Epub 2014 Apr 26.

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

Author information

1
Gastrointestinal Laboratory, Department of Small Animal Clinical Sciences, Texas A&M University, College Station, TX 77843, USA. Electronic address: tierarztbz@hotmail.com.
2
Gastrointestinal Laboratory, Department of Small Animal Clinical Sciences, Texas A&M University, College Station, TX 77843, USA.
3
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.

Abstract

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.

KEYWORDS:

Calprotectin; Cold-microwave technology; ELISA; Feces; Serum; Validation

PMID:
24780221
DOI:
10.1016/j.ab.2014.04.026
[Indexed for MEDLINE]

Supplemental Content

Full text links

Icon for Elsevier Science
Loading ...
Support Center