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Synthetic Multiepitope Protein-Based Plasmonic Immunosensor for Rapid, Label-Free Detection of Cryptococcus
Camila Erbereli1, Jaqueline Volpe2, Nicole Maia2
1Laboratório de Eletroquímica, Eletroanálise e Desenvolvimento de Sensores, Institute of Chemistry, State University of Campinas (UNICAMP), Campinas, SP 13083-970, Brazil.
Insights
This study developed a novel biosensor for cryptococcosis diagnosis. The surface plasmon resonance immunosensor uses a synthetic protein to detect fungal antibodies in human serum, improving diagnostic accuracy.
Area of Science:
- Mycology
- Immunology
- Biomedical Engineering
Background:
- Cryptococcosis is a severe fungal infection with high HIV-related mortality.
- Current diagnostic methods for cryptococcosis lack sensitivity, specificity, and point-of-care accessibility.
- Developing specific bioreceptors for detecting Cryptococcus antibodies is a significant challenge.
Purpose of the Study:
- To develop a novel biosensing approach for the early and accurate diagnosis of cryptococcosis.
- To evaluate a synthetic multiepitope protein as a bioreceptor for an SPR immunosensor.
- To establish a sensitive and specific method for detecting Cryptococcus antibodies in human serum.
Main Methods:
- Construction of a surface plasmon resonance (SPR) immunosensor.
- Utilizing a novel synthetic multiepitope protein (protein D) as the bioreceptor.
- Testing the SPR immunosensor with human serum samples for Cryptococcus antibody detection.
Main Results:
- The SPR immunosensor demonstrated limits of detection (LOD) of 0.1 μg mL⁻¹ and quantification (LOQ) of 0.5 μg mL⁻¹.
- The biosensor exhibited high selectivity and reproducibility in analyzing human sera.
- The system effectively discriminated between individuals with and without cryptococcosis.
Conclusions:
- A synthetic multiepitope protein is a viable bioreceptor for SPR immunosensors.
- The developed SPR immunosensor offers a promising tool for the serological diagnosis of cryptococcosis.
- This study presents the first plasmonic immunosensing system for detecting fungal Cryptococcus antibodies in human serum.
Abstract:
Cryptococcosis is a severe fungal infection, particularly in immunosuppressed individuals, causing over 112,000 HIV-related deaths annually. Early and accurate diagnosis is critical, but current methods often lack the necessary sensitivity, specificity, and accessibility for point-of-care use. A major challenge is identifying highly specific bioreceptors for detecting Cryptococcus-specific antibodies. This study addresses these diagnostic limitations by developing a novel biosensing approach. While biosensor technology holds significant promise for rapid, sensitive, and selective responses in healthcare, effective solutions for cryptococcosis, particularly antibody detection, remain challenging. The surface plasmon resonance (SPR) technique was employed as the transduction system for constructing the biosensor. A new synthetic multiepitope protein, called protein D, was evaluated as a bioreceptor for developing an SPR immunosensor. Protein D is a chimeric protein composed of five different peptides (H18, H21, H26, S4, and Hy49) linked in specific combinations. The proposed SPR immunosensor presented limits of detection (LOD) of 0.1 μg mL-1 and quantification (LOQ) of 0.5 μg mL-1. Analysis of human sera was performed with high selectivity and reproducibility, effectively discriminating between individuals with and without cryptococcosis. To date, no plasmonic immunosensing system has been reported for detecting fungal Cryptococcus antibodies in human serum. In brief, this study successfully demonstrated the viability of a synthetic multiepitope protein in an SPR immunosensor for the serological diagnosis of cryptococcosis.
