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A rapid enzyme-linked immunosorbent assay with two modes of detection for measuring cytokine concentration
Keishi Miwa1, Naoko Shibayama, Takeshi Moriguchi
1Toray Industries, Inc., New Frontiers Research Labs, Kamakura, Kanagawa, Japan. Keishi_Miwa@nts.toray.co.jp
Insights
A novel stick enzyme-linked immunosorbent assay (STICKELISA) system offers rapid, quantitative measurement of cytokines like Interleukin-6 (IL-6) and IL-8. This bedside method provides accurate results comparable to traditional plate ELISA, aiding critical care settings.
Area of Science:
- Biochemistry
- Immunology
- Medical Diagnostics
Background:
- Cytokine measurement is crucial in intensive care units (ICUs) for patient monitoring.
- Existing methods like plate ELISA can be time-consuming and require specialized laboratory settings.
- A need exists for rapid, point-of-care cytokine detection in critical care.
Purpose of the Study:
- To evaluate a novel polystyrene-based stick enzyme-linked immunosorbent assay (STICKELISA) system for measuring Interleukin-6 (IL-6) and Interleukin-8 (IL-8).
- To assess the system's speed, accuracy, and agreement with conventional plate ELISA.
- To determine the utility of STICKELISA for bedside cytokine quantification in intensive care patients.
Main Methods:
- Serum samples from 101 ICU patients were analyzed using the STICKELISA system.
- The system utilizes an immobilized-antibody ELISA stick and a noncontact spectrophotometer.
- Cytokine concentrations were determined both semi-quantitatively by visual color change and quantitatively via spectrophotometry.
Main Results:
- The STICKELISA spectrophotometric assay detected cytokines down to 100 pg/mL within 45 minutes.
- High agreement was observed between STICKELISA naked-eye observation and plate ELISA (94% for IL-6, 85% for IL-8).
- Strong correlation coefficients (0.88 for IL-6, 0.91 for IL-8) were found between STICKELISA spectrophotometric and plate ELISA determinations.
Conclusions:
- The STICKELISA system is a simple, rapid, and quantitative tool for bedside cytokine measurement.
- This method demonstrates good agreement with established plate ELISA techniques.
- STICKELISA holds significant potential for improving patient management in critical-care settings.
Abstract:
Interleukin (IL)-6 and IL-8 were measured in 101 serum samples collected from eight intensive-care unit patients using a polystyrene-based stick enzyme-linked immunosorbent assay (STICKELISA) system. This system consisted of an immobilized-antibody ELISA stick and a noncontact spectrophotometer. Cytokine concentration was detected by two ways: first, rapidly and semi-quantitatively by naked-eye observation of the color change and second, quantitatively using the spectrophotometer for accurate concentration determination. The spectrophotometric assay enabled the quantitation of as little as 100 pg/mL cytokine and took only 45 min to complete. There was a good agreement between the STICKELISA observations and data obtained using a plate ELISA system. The agreement between STICKELISA naked-eye observation and plate ELISA determination was 94 and 85% for IL-6 and IL-8, respectively. The correlation coefficients between the STICKELISA spectrophotometric determination and plate ELISA determination were 0.88 and 0.91 for IL-6 and IL-8, respectively, in a 0.1-5 ng/mL cytokine concentration range. These results demonstrate that the STICKELISA system is a simple, rapid, and quantitative method for bedside cytokine measurement in critical-care settings.
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