In the present study, monoclonal antibodies (MAbs) against recombinant histidine-rich protein (rHRP3) were developed and assessed for their potential in detection of Plasmodium falciparum HRP3. Hybridomas were obtained by fusion of Sp2/0 mouse myeloma cells and spleen cells from the mouse immunized with purified rHRP3. Three MAbs (IgG1 isotype) specific to rHRP3 were established and characterized by enzyme-linked immunosorbent assay (ELISA) and immunoblotting for sensitivity and specificity. Purification of MAbs from hybridoma cell culture supernatant and PAbs from rabbit anti-serum were carried out using Phenylpropylamine (PPA) HyperCel ^™ sorbent. The MAbs were able to detect rHRP3 and the HRP3 from P. falciparum spent medium. Sandwich ELISA was developed to quantify HRP3 in the spent medium of P. falciparum culture. The generated MAbs could be potentially used in immuno-based diagnostic systems for the detection of P. falciparum HRP.

Tuberculosis (TB) is a highly infectious disease caused by Mycobacterium tuberculosis. The World Health Organization (WHO) has reported more than 8 million new or relapse TB cases and 1.25 million deaths in 2023. Even though the global End TB strategy implemented by WHO has reduced the TB incidence rate between the years 2015 and 2023 up to 8.3%, TB remains one of the leading infectious causes of death worldwide. Active tuberculosis can affect any part of the body, and the clinical symptoms include fever, weight loss, and night sweats, whereas latent TB does not show any clinical symptoms. One of the significant concerns regarding the control of TB is the reservoir nature of latent tuberculosis. The Bacillus Calmette-Guerin (BCG) vaccine helps control TB and is still used globally to combat it. Currently, two methods are primarily used to identify tuberculosis infection: the Tuberculin Skin Test (TST) and the Interferon-Gamma Release Assay (IGRA). A significant disadvantage of these two methods is the inability to differentiate between latent and active tuberculosis. Accurate and timely diagnosis of TB is essential for effective management, mainly due to the emergence of multidrug-resistant strains. Specific biomarkers are required to evaluate tuberculosis infection, and novel biomarkers are needed to develop new diagnostic methods for tuberculosis. This article presents a brief review of recent developments in TB diagnostics, covering immunological, molecular, and monoclonal antibody-based platforms. The review also discusses the emerging role of artificial intelligence and deep machine learning platforms as complementary diagnostic tools.

The World Health Organization (WHO) has set forth a global call for eradicating malaria, caused majorly by the protozoan parasites Plasmodium falciparum and Plasmodium vivax. The lack of diagnostic biomarkers for P. vivax, especially those that differentiate the parasite from P. falciparum, significantly hinders P. vivax elimination. Here, we show that P. vivax tryptophan-rich antigen (PvTRAg) can be a diagnostic biomarker for diagnosing P. vivax in malaria patients. We report that polyclonal antibodies against purified PvTRAg protein show interactions with purified PvTRAg and native PvTRAg using Western blots and indirect enzyme-linked immunosorbent assay (ELISA). We also developed an antibody-antigen-based qualitative assay using biolayer interferometry (BLI) to detect vivax infection using plasma samples from patients with different febrile diseases and healthy controls. The polyclonal anti-PvTRAg antibodies were used to capture free native PvTRAg from the patient plasma samples using BLI, providing a new expansion range to make the assay quick, accurate, sensitive, and high-throughput. The data presented in this report provides a proof of concept for PvTRAg, a new antigen, for developing a diagnostic assay for P. vivax identification and differentiation from the rest of the Plasmodium species and, at a later stage, translating the BLI assay into affordable, point-of-care formats to make it more accessible.