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An ELISA-like time-resolved fluorescence immunoassay for microcystin detection
La-Mei Lei1, Ying-Song Wu, Nan-Qin Gan
1State Key Laboratory of Freshwater Ecology and Biotechnology, Institute of Hydrobiology, CAS, Donghu Nanlu No. 7, Wuchang, Wuhan 430072, PR China.
Clinica Chimica Acta; International Journal of Clinical Chemistry
|September 17, 2004
Summary
A new time-resolved fluorescence immunoassay (TRFIA) offers a sensitive and reliable method for detecting microcystins (MCs). This assay provides a broader detection range and improved sensitivity compared to traditional methods like ELISA.
Area of Science:
- Environmental Science
- Analytical Chemistry
- Biotechnology
Background:
- Development of a novel time-resolved fluorescence immunoassay (TRFIA) for microcystin detection.
- Utilizes anti-microcystin-LR (MCLR) monoclonal antibodies (MAbs) and a europium-labeled conjugate.
Purpose of the Study:
- To develop and validate a TRFIA for sensitive and reliable microcystin detection.
- To compare the performance of TRFIA with existing methods like ELISA and HPLC.
Main Methods:
- Preparation of anti-MCLR MAbs with cross-reactivity to related microcystins.
- TRFIA performed in an indirect competitive mode.
- Comparative analysis with indirect competitive enzyme-linked immunosorbent assay (ELISA) and high-performance liquid chromatography (HPLC).
Main Results:
- TRFIA demonstrated a wide quantitative detection range (0.01-10 ng/ml) and high sensitivity for MCLR.
- Achieved excellent reliability with low coefficients of variation (1.6-12.2%).
- Assay results for algal samples aligned with ELISA and HPLC, confirming practicality and reliability.
Conclusions:
- TRFIA presents a highly sensitive and reliable method for microcystin detection.
- It serves as a valuable alternative or substitute for conventional ELISA methods.
- The developed TRFIA is practical for detecting microcystins in environmental samples.