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A dipstick, dot-ELISA assay for the rapid and early detection of bladder cancer
A M Attallah1, H Helmi, E el-Helali
1Office of Biologic Research and Review, National Institutes of Health, Bethesda, MD.
Insights
A new immunoassay detects keratin in urine, offering a sensitive and specific method for early bladder cancer screening. This fast dot-ELISA requires minimal equipment and time, making it a promising tool for widespread use.
Area of Science:
- Biochemistry
- Oncology
- Immunology
Background:
- Bladder cancer diagnosis relies on invasive procedures.
- Need for sensitive, non-invasive early detection methods is critical.
- Keratin fragments in urine may indicate bladder cancer.
Purpose of the Study:
- To develop a sensitive and specific immunoassay for early bladder cancer detection.
- To create a rapid, user-friendly screening test using keratin detection.
- To evaluate the stability and correlation of the developed assay.
Main Methods:
- Development of a mouse monoclonal antibody (CK1K10) targeting keratin.
- Enzyme immunoassays (EIAs) for keratin detection in urine samples.
- Creation of a fast dot-enzyme-linked immunosorbent assay (dot-ELISA) on nitrocellulose.
Main Results:
- The CK1K10 antibody demonstrated high sensitivity and specificity for bladder cancer detection.
- The fast dot-ELISA showed high positive correlation with other EIAs.
- Urine-coated dipsticks maintained efficiency for over 5 days at room temperature and 3 months at -20°C.
Conclusions:
- The keratin dot-ELISA is a reliable and efficient screening test for early bladder cancer detection.
- The assay's simplicity and stability facilitate its use in various settings.
- This method offers a non-invasive approach to identifying potential bladder cancer cases.
Abstract:
A mouse monoclonal antibody (CK1K10) specific for keratin was developed. Enzyme immunoassays (EIAs) were performed to detect keratin in urine samples using CK1K10 and anti-mouse IgG. A high degree of sensitivity and specificity was obtained in the detection of bladder cancer patients. A fast dot-ELISA was developed in which urine is applied onto nitrocellulose (NC) sheets or dipsticks. Dipsticks coated with urine samples can be stored at room temperature for more than 5 days and at -20 degrees C for more than 3 months without loss of efficiency. A high positive correlation was found between the fast dot-ELISA and other EIAs. The assay does not require sophisticated equipment nor highly trained technical staff and can be performed in less than 30 min. The keratin dot-ELISA could be a reliable screening test for the early detection of bladder cancer.
