Diagnosis of Bacterial Meningitis and AMR Profile Using Molecular and Immunological Techniques

Abstract

Background: Bacterial meningitis (BM) is severe complication of central nervous system (CNS) and is often associated with high mortality and morbidity rates if not timely diagnosed and treated. Current diagnostic tools for BM and drug resistance suffer from lack of sensitivity due to paucibacillary nature of Cerebrospinal fluid (CSF). Objectives: The objective of the study is to develop rapid and efficacious immunological tools (Enzyme linked immunosorbent assay and Lateral flow) for diagnosis of BM in hospital settings. In addition, in-house molecular assays will be developed for diagnosis of extended spectrum beta lactamases and Carbapenems drug resistance in community and hospital acquired BM infection Methodology: A prospective observational study will be carried out in patients admitted in IPD awards of CIIMS, Nagpur. Diagnosis of BM will be done using conventional gold standards and by in-house designed nested polymerase chain reaction (PCR) for 8 etiological pathogens. An in-house ELISA assay will be standardized and optimized based on culture filtrates antigen isolated from clinical isolates of etiological pathogens associated with BM for immunological diagnosis. For diagnosis of drug resistance, in-house conventional PCR assay targeting ESBL (TEM, SHV and CTX-M) and Carbapenems (NDM, OXA-48, VIM) resistance genes will be standardized and optimized. The sensitivity and specificity of developed test will be determined by standard ROC curve using medical software. The diagnostic accuracy will be determined based on concordance with gold standards. Expected results: A simple, rapid ELISA assay will be developed for etiological diagnosis BM in hospital settings.  In addition, in house molecular assay will be developed for rapid identification of ESBL and Carbapenems drug resistance in BM cases. Conclusion: A simple, rapid, efficacious immunological and molecular tools for diagnosis of BM and drug resistance will be developed for improved management of cases in hospital settings as governance of bacterial meningitis in India is poor.