Dr. Promod Mehta

Dr. Promod Mehta |Clyto Access

Director, Centre for Biotechnology, Maharshi Dayanand University, Rohtak, India

Speaker

Expertise: Microbiology

Biography:

Dr Promod Mehta received his Ph.D. degree (Micobiology) from Panjab University, Chandigarh in 1988. He joined as a Lecturer (Microbiology) in 1989 in Delhi University College and continued till 2009 as Associate Professor (Microbiology). During his stayat the University of Delhi, he pursued postdoctoral research training at the Centers for Disease Control & Prevention (CDC), Atlanta, USA (1994-96) and University of Nebraska-Lincoln (UNL), NE, USA (1999-2001).

He has worked extensively in the area of TB pathogenesis including TB diagnostics for his postdoctoral research. Later, he also visited UNL, NE, USA as a Visiting Assistant Professor for six months each in 2003, 2005 and 2006.

Dr Mehta joined as a Professor at the Centre for Biotechnology (CBT), Maharshi Dayanand University, Rohtak in 2009 and is continuing as a Director, CBT since March, 2015. He has several projects funded by DBT, DST, ICMR and UGC and has focused his research on TB diagnostics as well as drug-designing. He has about 45 research papers in peer-reviewed journals and research proceedings.

Presentation:

Title: Immuno-PCR, a novel tool for an early diagnosis of tuberculosis

Abstract:

An early and accurate diagnosis of tuberculosis (TB) is important to initiate early anti-tubercular therapy so as to avoid unnecessary morbidity and mortality. Diagnosis of smear-positive pulmonary TB is considerably established, but the diagnosis of smear-negative PTB and the paucibacillary extrapulmonary TB exhibits serious challenges. We developed a novel immuno-polymerase chain reaction (I-PCR) assay for an early diagnosis of TB patients, which combines the versatility of enzyme-linked immunosorbent assay (ELISA) with the enormous amplification power of PCR. The coupling of reporter DNA with the detection antibody is a critical step, which was carried through streptavidin-biotin system or through a covalent bniding via a chemical linker succinimidyl 4-[Nmaleimidomethyl]-cyclohexane-1-carboxylate (SMCC). There were less incubation/wash steps with the use of SMCC thus leading to reduced background signals. We could detect up to 1 fg/mL of purified mycobacterial antigen 85B (Ag85B, Rv1886c) and ESAT-6 (Rv3875), which was ~ 105-106 lower than analogous ELISA. Detetion of Ag85B, ESAT-6 and cord factor (trehalose-6,6’ dimycolate) in body fluids of PTB and EPTB patients by I-PCR showed high sensitivity and specificity and that revealed superiority over ELISA. We also detected a cocktail of anti-ESAT-6, anti-Ag85B and anti-cord factor antibodies in sera of TB patients with promising results. However, detection of antigens in body fluids is considered more accurate in comparison to antibody detection as it is independent of host’s immune response. Therefore, we developed real-time I-PCR for the quantitative detection of mycobacterial PstS1 (Rv0934) in body fluids of TB patients with a wide dynamic range of 1ng/mL to 1 pg/mL that could facilitate monitoring the dynamics of disease and response to ATT at different clinical stages of TB patients. Attempts were made to further improve the accuracy of I-PCR and simplify the protocol for binding the reporter DNA and detection antibody through nanoparticles. The functionalized gold nanoparticles (GNPs) coupled with antibody and reporter DNA were characterized by transmission electron microscopic observations and PCR, while magnetic beads coupled with antibodies were characterized by magneto ELISA. We are presently focusing to analyze clinical TB samples by GNP based real-time I-PCR and magnetic beads coupled real-time I-PCR. Acknowledgement: This work was supported by a project funded by DBT (BT/PR8054/MED/29/710/2013), New Delhi.

Related Conferences :

International Biotechnology and Pharmaceutical Industry Forum