Rapid-format recombinant antibody-based methods for the diagnosis of Clostridioides difficile infection: Recent advances and perspectives

Abstract

Clostridioides difficile, the most common cause of nosocomial diarrhea, has been continuously reported as a worldwide problem in healthcare settings. Additionally, the emergence of hypervirulent strains of C. difficile has always been a critical concern and led to continuous efforts to develop more accurate diagnostic methods for detection of this recalcitrant pathogen. Currently, the diagnosis of C. difficile infection (CDI) is based on clinical manifestations and laboratory tests for detecting the bacterium and/or its toxins, which exhibit varied sensitivity and specificity. In this regard, development of rapid diagnostic techniques based on antibodies has demonstrated promising results in both research and clinical environments. Recently, application of recombinant antibody (rAb) technologies like phage display has provided a faster and more cost-effective approach for antibody production. The application of rAbs for developing ultrasensitive diagnostic tools ranging from immunoassays to immunosensors, has allowed the researchers to introduce new platforms with high sensitivity and specificity. Additionally, DNA encoding antibodies are directly accessible in these approaches, which enables the application of antibody engineering to increase their sensitivity and specificity. Here, we review the latest studies about the antibody-based ultrasensitive diagnostic platforms for detection of C. difficile bacteria, with an emphasis on rAb technologies.

Development and characterization of phage display-derived anti-toxin antibodies neutralizing TcdA and TcdB of Clostridioides difficile

“…However, considerable amount of research is still ongoing in this area ( 19 , 20 ). Among antibodies, recombinant antibody fragments, that is, fragment antigen binding (Fab), single-domain antibodies (sdAbs), and single-chain fragment variable (scFv) have received much attention in recent years ( 14 , 16 , 20 , 21 ). The Fab contains constant and variable regions of the heavy (C H1 and V H ) and light (C L1 and V L ) chains, which are connected by interchain disulfide bridges ( 12 ).…”

Clustering Disease of Clostridioides Difficile Infection: Implication for the Management in Internal Medicine

“…The clinical suspicion of CDI infection is related to the detection of the germ on the stool and the search for the toxins it produces in any case of watery diarrhea related to health care or in case of unexplained diarrhea [16,17]. Although numerous laboratory tests are available for the diagnosis of CDI, the optimal diagnostic strategy is still debated given the differences in sensitivity and specificity of the various methods.…”

Transient expression of anti-HrpE scFv antibody reduces the hypersensitive response in non-host plant against bacterial phytopathogen Xanthomonas citri subsp. citri

“…Recombinant-antibody-mediated resistance in plants was first reported in the early 1990s 16 , and since then, different recombinant antibodies, e.g., fragment antigen-binding region (Fab), single chain variable fragments (scFvs), and single-domain antibody (sdAbs), have been expressed in plants against different plant pathogens 17 – 19 . Among the various forms of recombinant antibodies, scFv is the most applicable type, which retains antigen specificity and function of the full-size antibody 20 . Phage display is the most conducive technology for the development of recombinant scFv fragments.…”