The Division of Biochemistry and Biotechnology is actively involved in disease diagnosis during epidemics and outbreak, operational research, manpower development, advisory role and other multifarious activities towards prevention and control of a cascade of epidemic-prone disease of larger public health importance.The division has two major infra structural units:

Biotechnology Wing

• Microbial PCR & DNA Fingerprinting Laboratory

• Molecular Diagnostics & Gene Cloning Laboratory

Biochemistry Wing

• Toxicology & Environmental Biochemistry Laboratory

• Clinical Biochemistry Laboratory

• IDD Laboratory

Objectives of Biotechnology Wing

• Molecular Diagnosis & Molecular Epidemiology of Epidemic-prone diseases.

• DNA Fingerprinting / Gene Sequencing of Pathogens.

• PCR/RT-PCR/DNA Hybridization for ultimate diagnosis.

• Tracking-down the source of infection of emerging & re-emerging diseases.

• Molecular differentiation of strains, detection of virulent/ drug resistant and new strains.

• Sub-typing of strains based on genotypes.

• Indigeneous synthesis of PCR primers and probes of important diseases.

Objectives of Biochemistry Wing

• Assay of iodine in salt / urine, thyroid function test and training for IDD Monitoring under National Iodine Deficiency Disorders Control Programme (NIDDCP).

• Studies on heavy metal and pesticides poisoning, chemical analysis of water, particularly anions and cations of public health importance.

• Detection of sangunarine toxicity in clinical cases of epidemic dropsy using highly sensitive HPLC method.

• Studies on anti-microbial resistance profile in clinical isolates of Klebsiella pneumoniae

• Studies on drug resistance gene(s) in M.tuberculosis and klebsiella. 

• Impart specialised referral services and support to outbreak investigations for providing supportive data related to haematology and biochemistry profiles on clinical specimens collected during the investigations and outbreaks.

Main achievements 

• Developed completely indigenous facility for automated gene sequencing of wild polio virus type 1, Dengue, HIV, Viral Hepatitis (HBV)

• Complete nucleotide sequence of 6 kb mini-circle gene of P.virak mitochondrial DNA has been successfully carried out.

• Nucleotide sequences of envelope region of dengue virus have been sequenced.

• Strain differentiation of P.falciparum by PCR using known variable gene makers (MSA-1, MSA-2, TRAP & KAHRP) has been standardised.

• Development of PCR for detection of mecA gene in Staphylococcus aureus.

• Molecular differentiation of drug resistant strains of K.pneumoniae.

• Detection of rifampicin resistance in Mycobacterium tuberculosis by identifying mutations in rpoB gene and PCR based diagnosis of Mycobacterium tuberculosis.

• Isolated and charecterised Histone 2B gene of P.vivax

• Development of liquid chromatographic method to assay tetracycline in serum samples

• Development of Fluorescence detection method for estimating sanguinarine using reverse phase high performance liquid chromatography in serum of suspected epidemic dropsy patients as well as in mustard oil

• As a National Reference Centre for IDD monitoring, this division has contributed immensely to NIDDCP by imparting extensive training to programme associated personnels from almost all States and UTs of India. This is also one of the four National Reference Centres which has developed expertise in Urinary Iodine Estimation and Thyroid Function Tests for evaluation of field surveys

• Standardised the PCR based diagnosis of P.vivax/P.falciparum from filter paper dried blood spots and stained blood smear. 

Significance of Molecular biology/ Biotechnology Set-up at NICD

Microbiological culture and conventional characterisation of disease pathogens are virtually replaced by current day PCR and gene sequence studies intended to provide more precise identity in ultimate terms of DNA fingerprints. In recent years, diagnostic molecular biology and molecular epidemiology has evolved from mere technology of curiosity into a field of vast diversity of diagnosis, epidemiology, therapeutics and vaccine development and in fact, the beginning of the millennium 2000 is bound to be the era of molecular medicine, The institution has established one of the finest application-oriented Biotechnology and Molecular biology set-up for molecular characterisation of epidemic-prone diseases in South East Asia. The laboratory, besides having highly skilled manpower and all basic facilities, is equipped with latest state of the art instrumenatation facilities.

Molecular Characterisation: PCR and DNA Fingerprinting of Disease pathogens

Molecular characterisation and molecular epidemiology of disease pathogens using Automated DNA sequencer ABI Prism 373 for DNA fingerprinting, DNA/RNA Synthesiser for primers and probes synthesis, Robotic Molecular Biology Lab Station (Catalyst-800) for genetic manipulation and PCR Thermal Cycler for PCR/RT-PCR. The set-up provides opportunity to study the molecular epidemiology of diseases to trace down the origin and source of an infection and to develop better disease surveillance mechanism in the country, particularly for emerging and re-emerging diseases. Fool proof facility for Molecular characterisation of wild Polio virus isolates. Dengue, HIV, Viral hepatitis (HBV & HCV), Vivax & falciparum malaria, Anthrax, Enteroviruses etc. have already been established.

Molecular Diagnostics & Gene Cloning Laboratory

i) Nucleotide sequence analysis of some of the expressed genes of P.vivax by screening of DNA gene library have been achieved. This will help in providing useful data to understand better the basic malaria biology and to identify P.vivax genes which may have potential for new future targets for drug designing and therapeuties, for vaccine candidates, for drug resistant markers, for species specific diagnosis etc.

ii) Methods for gene cloning, sub-cloning and protein expression established in the process of isolation and partial characterisation of Histone 2B gene of P.vivax.

iii) Sequence analysis of 6 Kb mini circle gene of P.vivax completed. It will be of great help in critical molecular characterisation of drug resistant cases and may also have therapeutic uses, (since mitochondria is a power-house and genes encoded by the mini circle are expressed at erythrocytic stages and exoerythrocytic stages). These studies may have important role in future malaria control programme.

iv) Maintenance of vectors and cells for Gene Cloning work.

Indigenous synthesis of PCR primers for important diseases

Polio/ and enteroviruses (EV70, EV71), Dengue, Hepatitis B/C, HIV, Anthrax, Plague, O139 and 01 V. cholerae, Rabies, JE, P.vivax, P.falciparum, HSV, Measeles, Paramixovirus, Nipha virus, Mycoplasmia, Legionella phneumophilla, M.tuberculosis, K. pneumoniae, Filaria, M-13, T3/T7, SP6 universal primers etc.

 Research Projects :

1. Molecular Epidemiology of Wild Poliovirus isolates

   1. Molecular epidemiology of wild Poliovirus isolates using RT-PCR and gene sequencing

   2. Strains classification into various genotypes

   3. Study to establish epidemiological linkages within wild poliovirus type 1,2,3 isolates

   
   

2. Molecular sub-typing of HBV by partial genome sequencing of HBs gene

   1. Sub-typing of HBV by PCR/Sequencing of 259 bp HBsAg gene encompassing codon 122 (d/y) and codon 160 (r/w) in serological confirmed cases from different hospitals.

   2. Sub-typing of HBsAg positive cases from 12 identified states of India under the collaborative Surveillance Project with the Division of Epidemiology and Microbiology.

3. Molecular characterisation of Dengue virus Type-2 isolates will be carried out to identify strain differentiation and mutations in the Pre-M, M and Env/NSI gene of region of the virus isolates from Dengue/ DHF outbreaks

   1. RT-PCR/ Cloning of C-PrM and Env-NSI gene junction in suitable Vector(s) vis-à-vis direct amplification of the said genes directly from confirmed isolates using group/quasi-species specific primers.

   2. Gene sequencing of the amplified C-PrM and Env-NSI genes using D-rhodamine based Big-dye primer/Big-dye terminator chemistry on retrospective and prospective clinical isolates from India and other regions of South-East Asia.

   3. Gene sequence comparison and analysis of the deduced phylogenic tree to ascertain epidemiological linkage and molecular evolution of Dengue (DF) and Dengue haemorrhagic fever (DHF) in the Indian sub-continent.

   4. Molecular detection of dengue virus in the field caught naturally infected Aedes mosquitoes to serve as an early warning signal for possible dengue outbreak.

4. Molecular Typing of Human Immunodeficiency Virus 1 Strains Circulating in India.

   1. Molecular diagnosis and epidemiology of HIV using PCR based methods.

   2. Gene sequencing of full length / short segments of gag and env genes for sub-typing and strain differentiation of HIV 1 subtype ‘C’ circulating in India.

   3. Study of drug resistance mutations in the reverse transciptase and proteas genes of HIV - 1 in selected clinical isolates.

5. Molecular diagnosis and characterisation of plasmodium              

   1. Molecular diagnosis of Plasmodia Species from stained blood smears and filter paper dried blood spot.

   2. PCR based strain differentiation in Plasmodium falciparum

   3. Sequence analysis of 6 kb mini circular mitochondrial genome of P.vivax and P.falciparum Indian isolates.

6. Molecular confirmation of suspected isolates of B.anthracis by PCR and Gene sequencing of S-layer and pX01/pX02 virulence plasmid genes.

   
   
    

7. Molecular charecterisation of drug resistance in K.pneumoniae: based  on ESBL production and plasmid profile.

8. Rifampicin- resistant mutations in rpoB gene for strain differentiation of Multi Drug Resistant Tuberculosis

9. National Reference Laboratory for IDD Monitoring supporting the NIDDCP by providing laboratory services for monitoring levels of iodine in salt samples, urine samples and thyroid hormones.

10. To monitor the levels of anions, cations and heavy metals in drinking water.

11. Floride and Fluorosis; Analysis of water samples

 

Manpower Development/Training Courses, Seminars, Symposia, Workshops

The division is actively involved in the activities of manpower development, training courses, conferences, workshops, seminars and symposia organised by the Institute from time to time to support the National Health Programmes.

The division is regularly organizing training cources for Programme Officer and Technical Staff of various States and UTs working under Iodine Deficiency Disorder Control Programme.

The division is also organising Trainers Training Workshop for medical professionals and public health engineers for fluorosis prevention and control measures.

Outbreak Investigations

The division is providing RT-PCR based molecular diagnostic laboratory support for investigation and confirmation during various outbreak of known and unknown aetiology.

Clinical Biochemistry - Providing a lead in outbreak investigations for Liver Function Test, Kidney Function Test, Blood Sugar, Lipid Profileetc.

Participation in Conferences, Workshops, Seminars & Symposia

Faculty members of the division are participating in Conferences, Wrokshops, Seminars/Symposia. The division is also publishing scientific findings in National and International Journals. 

Membership of Expert Committees/ Advisory Panel/ Honours 

The faculty members are on the panels of various Scientific Expert Committee/Advisory Board. Faculty members are also acting as Doctoral Committee members Guide and Examiners for Ph.D/M.Sc couces of various Indian Universities.