Surface plasmon resonance-based inhibition assay for real-time detection of Cryptosporidium parvum oocyst
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Chang Duk Kang1, Cuong Cao, Jeewon Lee
1School of Chemical and Biological Engineering, Seoul National University, Seoul 151-744, Republic of Korea.
Abstract:
A surface plasmon resonance (SPR)-based inhibition assay method using a polyclonal anti-mouse IgM arrayed Cryptosporidium sensor chip was developed for the real-time detection of Cryptosporidium parvum oocysts. The Cryptosporidium sensor chip was fabricated by subsequent immobilization of streptavidin and polyclonal anti-mouse IgM (secondary antibody) onto heterogeneous self-assembled monolayers (SAMs). The assay consisted of the immunoreaction step between monoclonal anti-C. parvum oocyst (primary antibody) and oocysts, followed by the binding step of the unbound primary antibody onto the secondary antibody surface. It enhanced not only the immunoreaction yield of the oocysts by batch reaction but also the accessibility of analytes to the chip surface by antibody-antibody interaction. Furthermore, the use of optimum concentration of the primary antibody maximized its binding response on the chip. An inversely linear calibration curve for the oocyst concentration versus SPR signal was obtained in the range of 1x10(6)-1x10(2)oocystsml(-1). The oocyst detection was also successfully achieved in natural water systems. These results indicate that the SPR-based inhibition assay using the Cryptosporidium sensor chip has high application potential for the real-time analysis of C. parvum oocyst in laboratory and field water monitoring.
Insights
A new surface plasmon resonance (SPR) method detects Cryptosporidium parvum oocysts in real-time. This assay uses a specialized sensor chip for accurate water monitoring.
Area of Science:
- Environmental Science
- Analytical Chemistry
- Biotechnology
Background:
- Cryptosporidium parvum oocysts pose a significant threat to water safety.
- Accurate and real-time detection methods are crucial for public health.
- Existing detection methods may lack sensitivity or speed.
Purpose of the Study:
- To develop a novel surface plasmon resonance (SPR)-based inhibition assay for rapid Cryptosporidium parvum oocyst detection.
- To create and optimize a Cryptosporidium sensor chip for enhanced assay performance.
- To evaluate the assay's effectiveness in detecting oocysts in natural water samples.
Main Methods:
- Fabrication of a Cryptosporidium sensor chip via immobilization of streptavidin and anti-mouse IgM on self-assembled monolayers (SAMs).
- Development of an SPR-based inhibition assay involving immunoreaction between anti-C. parvum oocyst antibody and oocysts.
- Optimization of primary antibody concentration and utilization of secondary antibody for enhanced binding.
Main Results:
- An inversely linear calibration curve was established for oocyst concentrations from 1x10^6 to 1x10^2 oocysts/mL.
- The SPR assay demonstrated successful real-time detection of Cryptosporidium parvum oocysts.
- Effective oocyst detection was achieved in natural water systems, validating field applicability.
Conclusions:
- The developed SPR-based inhibition assay offers a highly sensitive and real-time method for Cryptosporidium oocyst detection.
- The specialized sensor chip enhances assay performance through improved immunoreaction yield and analyte accessibility.
- This method shows significant potential for both laboratory and field-based water quality monitoring.
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