Liquid Redox Probe-Free Plastic Antibody Development for Malaria Biomarker Recognition.

Malaria is a major public health challenge worldwide and requires accurate and efficient diagnostic methods. Traditional diagnostic approaches based on antigen-antibody interactions are associated with ethical and economic concerns. Molecularly imprinted polymers (MIPs) offer a promising alternative by providing a complementary polymer structure capable of selectively binding target molecules. In this study, we developed a liquid, redox-probe-free, MIP-based electrochemical biosensor to detect the Plasmodium falciparum malaria marker histidine-rich protein (HRP2) at the point-of-care (PoC). The imprinting phase consists of the electropolymerization of the monomer methylene blue (MB) in the presence of the target protein HRP2 at the working electrode (WE) of the modified carbon screen printed electrode (C-SPE). Subsequent removal of the protein with proteinase K and oxalic acid yielded the MIP material. The sensor assembly was monitored by cyclic voltammetry (CV), Raman spectroscopy and scanning electron microscopy (SEM). The analytical performance of the biosensor was evaluated by square-wave voltammetry (SWV) using calibration curves in buffer and serum with a detection limit of 0.43 ± 0.026 pg mL-1. Selectivity studies showed minimal interference, indicating a highly selective assay. Overall, our approach to detect the HRP2 infection marker offers simplicity, cost-effectiveness and reliability. In particular, the absence of a redox solution simplifies detection, as the polymer itself is electroactive and exhibits oxidation and reduction peaks.

Diagnostic applications of microsphere-based flow cytometry: A review.

Microsphere-based flow cytometry is a highly sensitive emerging technology for specific detection and clinical analysis of antigens, antibodies, and nucleic acids of interest. In this review, studies that focused on the application of flow cytometry as a viable alternative for the investigation of infectious diseases were analyzed. Many of the studies involve research aimed at epidemiological surveillance, vaccine candidates and early diagnosis, non-infectious diseases, specifically cancer, and emphasize the simultaneous detection of biomarkers for early diagnosis, with accurate results in a non-invasive approach. The possibility of carrying out multiplexed assays affords this technique high versatility and performance, which is evidenced in a series of clinical studies that have verified the ability to detect several molecules in low concentrations and with minimal sample volume. As such, we demonstrate that microsphere-based flow cytometry presents itself as a promising technique that can be adopted as a fundamental element in the development of new diagnostic methods for a number of diseases.

Electrochemical immunosensor for detection of Plasmodium vivax lactate dehydrogenase.

BACKGROUND: Malaria is a disease that affects many tropical and subtropical countries, including Brazil. The use of tests for malaria detection is one of the fundamental strategies recommended by the World Health Organization for the control and eradication of the disease. The lack of diagnostic tests leads to an increase in transmission and non-reporting cases. OBJECTIVES: This work described an electrochemical immunosensor for detecting Plasmodium vivax lactate dehydrogenase antigen (Ag-PvLDH). METHODS: The device has developed by immobilising egg yolk IgY antibodies (Ab-PvLDH) on a gold electrode surface using cysteamine as linker. The immunosensor fabrication was followed by differential pulse voltammetry, and contact angle measurements were performed to characterise the modified gold electrode surface. FINDINGS: The results for Ag-PvLDH determination exhibit a linear response at 10-50 µg mL-1 concentration range, with a limit of detection of 455 ng mL-1. The excellent selectivity of the device was confirmed. MAIN CONCLUSIONS: The developed immunosensor showed a good performance, therefore, it can be considered an alternative test to detect malaria caused by P. vivax.

Bacillus subtilis spores as delivery system for nasal Plasmodium falciparum circumsporozoite surface protein immunization in a murine model.

Malaria remains a widespread public health problem in tropical and subtropical regions around the world, and there is still no vaccine available for full protection. In recent years, it has been observed that spores of Bacillus subtillis can act as a vaccine carrier and adjuvant, promoting an elevated humoral response after co-administration with antigens either coupled or integrated to their surface. In our study, B. subtillis spores from the KO7 strain were used to couple the recombinant CSP protein of P. falciparum (rPfCSP), and the nasal humoral-induced immune response in Balb/C mice was evaluated. Our results demonstrate that the spores coupled to rPfCSP increase the immunogenicity of the antigen, which induces high levels of serum IgG, and with balanced Th1/Th2 immune response, being detected antibodies in serum samples for 250 days. Therefore, the use of B. subtilis spores appears to be promising for use as an adjuvant in a vaccine formulation.

Electrochemical immunosensor for detection of Plasmodium vivax lactate dehydrogenase

BACKGROUND Malaria is a disease that affects many tropical and subtropical countries, including Brazil. The use of tests for malaria detection is one of the fundamental strategies recommended by the World Health Organization for the control and eradication of the disease. The lack of diagnostic tests leads to an increase in transmission and non-reporting cases. OBJECTIVES This work described an electrochemical immunosensor for detecting Plasmodium vivax lactate dehydrogenase antigen (Ag-PvLDH). METHODS The device has developed by immobilising egg yolk IgY antibodies (Ab-PvLDH) on a gold electrode surface using cysteamine as linker. The immunosensor fabrication was followed by differential pulse voltammetry, and contact angle measurements were performed to characterise the modified gold electrode surface. FINDINGS The results for Ag-PvLDH determination exhibit a linear response at 10-50 µg mL-1 concentration range, with a limit of detection of 455 ng mL-1. The excellent selectivity of the device was confirmed. MAIN CONCLUSIONS The developed immunosensor showed a good performance, therefore, it can be considered an alternative test to detect malaria caused by P. vivax.