Development of Affordable Methods and Protocols for Diagnosis of Clostridium difficile Infection

Abstract

Background: Clostridium difficile infection (CDI) is a major emerging pathogen in India. Its diagnosis is crucial and gradually becoming primary demand of the healthcare system. The existing diagnostics facility provides multiple pathological techniques like Enzyme linked immunosorbent assay (ELISA), Lateral flow, Nucleic acid amplification test (NAAT), etc. which are considered as the most reliable one but due to its labour-intensive, time-consuming and complicated procedure, it has a longer turnaround time and the results are not satisfactory.

Objective: To develop and Evaluate Immunological & Molecular assays using advance proteomics targeting novel biomarkers for diagnosis of CDI.

Methodology: The present study will focus on using advance proteomics for characterization of proteins. This will be followed by peptide designing by the Kolaskar model &antipeptides synthesis. The identified peptides/lysate will be used to develop ELISA. For molecular method, Multiplex PCR will be developed which will reduce the chance of a false-negativity.

Expected Results: We propose to use the stool & culture secretome, a cocktail of several bacterial proteins, with broad specific immune response effective for diagnosis. Such toxigenic proteins may be detected using simple ELISA based tests, which is rapid yet affordable and even have scale up capacity to be converted into rapid point of care test in the form of lateral flows. Such assays, if developed, can aid clinicians in bedside screening for CDI and initiate specific treatment in suspected culture-negative cases to avoid complications associated with empirical regimes.

Conclusion: Our study will be intended to identify novel biomarkers specific to CDI develop a rapid & affordable, point of care assay which can be easily performed in any clinical laboratory, which will ultimately be helpful in the early definitive diagnosis of CDI, which if untreated will burst out as a major health problem in developing countries.