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  • Wednesday | 24-July-2019

Molecular Medicine

Molecular Medicine

Dr Paras Singh Head of the Department View Profile
     

Research Interests:

  • Identification of molecular markers associated with tuberculosis;
  • Lung cancer;
  • HIV;
  • Rapid molecular detection of Mycobacterium tuberculosis in human clinical specimens.

 List of Equipments:

  • PCR
  • COBAS TaqMan 48
  • Refrigerated centrifuge
  • Class II Type B2 Biohazard Safety Cabinet
  • Laminar Air Flow
  • FACS Count
  • Deep Freezers
  •  Gel electrophoresis
  •  NanoDrop
  • Gel-Doc

 Research and Diagnostic Activities:

 Tuberculosis (TB) continues to be a major health burden; being one of the top 10 causes of mortality worldwide.  The high rate of incidence of tuberculosis in India indicates a genetic predisposition to susceptibility to the disease. Our Department provides diagnostic services for PTB and EPTB by TB-PCR and RT-PCR. The Dept. of Molecular Medicine also determines CD3/CD4 and CD8 counts in clinical specimens by BD-FACS in healthy and TB cases. We are also engaged in identification of gene polymorphisms in Indian population that may alter susceptibility to pulmonary or extra-pulmonary M. tuberculosis infection. The polymorphisms are being studied at both nucleic acid and protein levels. We have identified genetic variants in different human genes, such as, CCR2, CCL2, TLR4, etc.  The CCL2 (−2518A/G) and TLR4 (D299G or 896A/G & T399I or 1196C/T) gene polymorphisms were studied by PCR-RFLP method, while the CCR2 (V64I or 190G/A) gene polymorphism was done by ARMS-PCR method.  Our findings indicated that the variant alleles of genes CCR2 (190 G to A) and TLR4 (896A/G & 1196C/T) make Indian population vulnerable towards developing tuberculosis. Further analysis with larger sample sizes is being done in the Department. The outcome from this research work may allow application of these biomarkers for better management of tuberculosis in Indian populaion.

 The Department of Molecular Medicine is also working in collaboration with the Department of Biochemistry, University of Delhi for a collaborative project entitled “Identification of mycobacterial proteins and novel antigenic epitopes having immunodiagnostic potential and development of reagents for point of care test for tuberculosis”.  The Department is also working towards molecular profiling of ‘driver mutations’ in non-small cell lung carcinoma (NSCLC). We are screening the formalin-fixed paraffin-embedded tissue biopsies for EGFR (Exon 18-21) mutations and EML4-ALK fusions. Simultaneously, we are also optimizing protocols for in-house detection of other mutations in the tissue specimens. The identification of such mutations is significant for defining and monitoring the treatment outcomes of the NSCLC patients. Another research project entitled “Evaluation of in-house Loop-Mediated Isothermal Amplification (LAMP) assay for rapid diagnosis of Mycobacterium Tuberculosis” is ongoing in our Department. This research work aims to develop a rapid, cheaper yet efficient molecular diagnostic assay for tuberculosis in humans.

 The Department submitted 4 research proposal concept notes in ICMR to avail extra-mural funding entitled 1) “Comparative evaluation of loop-mediated isothermal amplification assay and cartridge-based nucleic acid amplification for diagnosis of smear negative M. tuberculosis cases”;  2) “Molecular profiling of non-small cell lung cancer tumor specimens to aid in anti-cancer treatment for better response in patients”;  3) “Co-relation of distinct cytokine signatures with different forms of tuberculosis infection”;  4) “Identification of miRNA signatures in tuberculosis infection”.

The major focus of the Departmental research activities is identification of biomarkers for PTB, EPTB and lung cancer. We are currently looking for differentiating biomarkers, such as, chemokines or their receptors and ligands, different gene variants (EGFR, XRCC4), other SNPs (CCR2, CCR5, MCP-1, ICAM, TNF, IFN-γ, ESAT6, DC-SIGN, IP10, TLR4 and TLR9 etc. for diseases, such as, TB, HIV, lung cancer. Since its establishment, the Department has published 9 research articles in peer-reviewed international journals, such as, Infection, Genetics & Evolution, Journal of Microbiological Methods, Meta Gene, etc.

 

The Department received 11 applications for undergoing 3-months training on “Molecular Biology Techniques”. Of these 11, the Department short-listed 2 applications for the training.

 With increasing demand for better services to TB patients, our efforts will be continuously introducing latest molecular tools for identification of Mtb in clinical specimens, with major focus on early diagnosis and treatment of tuberculosis in pediatric cases. We also aim to develop strategies for efficient molecular diagnosis of MDR/XDR cases of tuberculosis.

 Recent Publications:

 1.                  Singh P, Rajput R, Mehra NK, Vajpayee M, Sarin R. HLA variations determine genetic predisposition to HIV-1 Infection: A Study in Indian population. Journal of Genetics. 2019. Under review.

2.                  Singh P, Rajput R, Mehra NK, Vajpayee M, Sarin R. Cytokine gene polymorphisms among North Indians: Implications for genetic predisposition? Infect Genet Evol. 2019 pii: S1567-1348(19)30105-4.

3.                  Rajput R, Singh P, Sarin R, Sethi P, Sharma S. Diagnostic accuracy of loop-mediated isothermal amplification assay for extra-pulmonary tuberculosis in Indian population. Journal of Microbiological Methods.2019 158:59-65.

4.                  Singh P, Rajput R, Sarin R, Tayal D. “Significance of CCL2, CCR2 and TLR4 polymorphisms (896 A/G and 1196 C/T) in tuberculosis risk in Indian population”. Meta Gene 2018; 18:184-190.

5.                  Verma AKSarin RArora VKKumar GArora JSingh PMyneedu VP. Amplification of Hsp 65 gene and usage of restriction endonuclease for identification of non-tuberculous rapid grower mycobacterium. Indian J Tuberc. 2018; 65(1):57-62.

6.                  Singh P, Singh M, Bhalla M, Singhla R, Gumma PK, Tayal D, Myneedu VP, Sarin R. “Synergetic effect of TLR4 gene (D299G and T399I) polymorphisms in pulmonary tuberculosis susceptibility. Universal Journal of Medical Science 2015;3(1): 19-27. http://www.hrpub.org DOI: 10.13189/ujmsj.2015.030103.

7.                  Verma AK, Kumar G, Arora J, Singh P, Arora VK, Myneedu VP, Sarin R. Identification of mycobacterial species by PCR restriction enzyme analysis of the hsp65 gene - an Indian experience.  Can J Microbiol. 2015;61(4):293-6. doi: 10.1139/cjm-2014-0525. Epub 2015 Jan 21.

8.                  Singh P, Singh M, Tayal D, Myneedu VP, Bhalla M, Adlakha P, Sarin R. The Clinical Utility of Polymerase Chain Reaction and Adenosine deaminase (ADA), for the Diagnosis of Pleural Tuberculosis: Indian Scenario. Immunology and Infectious Diseases 2014; 2(2): 13-21.

9.                  Singh M, Singh P, Myneedu VP, Bhalla M,  Adlakha P, Verma P,  Sarin R.  “Evaluation of Polymerase Chain Reaction and CobasTaqMan Real Time PCR in the Diagnosis of Tuberculosis: Indian Prospective. Immunology and Infectious Diseases 2013;1(1): 1-9.

 

10.              Singh P, Singh M, Adlakha P, Verma P, Myneedu V P, Sarin R. Comparative Evaluation of PCR with Commercial Multiplex M. tuberculosis Detection Kit. Immunology and Infectious Diseases 2013; (2): 19-26.

 

11.              Mehra NK, Singh P, Singh M, Najmi N, Kaur G, Sharma SK, Katoch K, Katoch VM - Shared and Distinct Variants of MHC and non MHC genes in pulmonary TB and leprosy NIH publisher 2009. In 44th US-Japan cooperative Medical Science Program, Forty fourth Tuberculosis and Leprosy Research Conference, Fukoka, Japan, p145-149, July 2009.

12.              Singh P, Kaur G, Sharma G, Mehra NK. Immunogenetic basis of HIV-1 infection, transmission and disease progression. Vaccine. 2008;26(24):2966-80. Epub 2008 Feb 4.

13.              Kaur G, Singh P, Rapthap CC, Kumar N, Vajpayee M, Sharma SK, Wanchu A, Mehra NK. Polymorphism in the CCR5 gene Promoter and HIV-1 infection in North Indians. Human Immunol. 2007;68(5):454-461. 

14.              Kaur G, Singh P, Rapthap CC, Kumar N, Vajpayee M, Sharma SK, Wanchu A, Mehra NK. Distribution of CCR2 polymorphism in HIV-1 infected and healthy subjects in North India. Internat J Immunogenet. 2007;34(3):153-156.

15.              P Singh, G.Sharma, G.Kaur, M.Vajpayee, S Sharma, N.Mehra. Host immunogenetics and HIV-1 infection (O-16) Tissue Antigens 2008;72, 227–313.

16.              Panigrahi A, Shiddqui J, Bhowmik D, Singh P, Tiwari S, Guleria S. Neutrophil gelatinase-associated lipocalin (NGAL) as a biomarker to predict acute tubular necrosis (ATN) in renal transplant allografts (P-O16) Tissue Antigens 2008;72, 227–313.