Background: bacterial pathogens and their toxins present analytical challenges for rapid and specific detection, contributing to over 600 million cases of illness annually and worsening antimicrobial resistance (AMR). Conventional detection methods are useful but limited. Single-domain antibodies (sdAbs) offer alternative recognition elements with unique biochemical and engineering benefits, enabling the development of nanobody-based immunoassays and biosensing platforms that provide fast, highly selective, and reliable detection of bacterial pathogens and toxins in both food and clinical environments. Objectives: this systematic review assesses the analytical and functional performance of nanobody-based immunoassays and sensing formats for detecting bacteria and toxins across food and clinical samples. Methods: following PRISMA guidelines, major scientific databases were used to gather research, resulting in 32 eligible studies published between 2011 and 2025. Results: data collected included assay platforms, target bacteria and toxins, limit of detection, sensitivity, specificity, matrix recovery, and practicality. Risk of bias was evaluated using an adapted QUADAS-2 framework. The review shows that nanobody-based immunoassays have achieved high performance, thermostability, compatibility with genetic engineering, and versatile assay design. When combined with advanced transduction and signal amplification strategies, these systems contribute to the development of highly sensitive and user-friendly bioanalytical platforms for detecting bacteria and toxins. Conclusions: however, most studies relied on spiked samples and lacked large-scale validation, emphasizing the need for standardized benchmarking and real-world testing.

Since 2020, SARS-CoV-2 viral culture has been a pivotal experimental tool for elucidating viral biology and supporting translational research. This study provides a five-year (2020–2025) retrospective and bibliometric mapping of 4405 unique publications retrieved from Scopus, Web of Science, and PubMed. Quantitative science mapping was integrated with a qualitative retrospective assessment of the ten most cited articles to identify influential biological and technological landmarks. The bibliometric results show a publication peak in 2021, with the United States and China as leading contributors. Thematic evolution shifted from foundational viral isolation to variant-specific investigations (Delta and Omicron) and therapeutic development. Retrospective analysis highlights a significant technological transition from standard cell lines to engineered models, including 3D organoids and deep mutational scanning approaches for the characterization of immune escape and viral fitness. Importantly, manual verification identified author name ambiguity as a major methodological limitation distorting productivity metrics. Overall, viral culture remains a critical bridge for validating phenotypic traits and establishing infectivity windows, providing a data-driven framework for future pandemic preparedness.