Major Research Achievements

Prophylactics

Viral vectored vaccines

1. Glycoprotein E (gE) deleted bovine herpesvirus-1 has been developed and evaluated as IBR marker vaccine candidate.
2. Rescue of Newcastle virus strain F with VP2 gene of infectious bursal disease virus of poultry has been established.
3. Developed and evaluated the recombinant NDV vaccine strain R2B with avirulent fusion protein cleavage site in chickens.

DNA vaccines

1. A RNA replicon based DNA vaccine vector for enhanced heterologous gene expression in mammalian cells was developed and evaluated to express reporter genes (e.g. Green Fluorescent Protein (GFP) and β-galactosidase) and protective immunogenic genes (e.g. rabies glycoprotein, canine parvovirus VP2).
2. A bicistronic DNA vector encoding HN and F gene of Newcastle disease virus was constructed.
3. Construction of bicistronic DNA vector encoding 35 kDa, 16.8 kDa (thiol peroxidase) and 10 kDa PE proteins of Mycobacterium avium paratuberculosis individually along with IFNγ and study of their immunogenicity in mouse model.
4. DNA constructs encoding canine mammary tumor associated antigens developed for CMT prophylaxis.
5. DNA vaccine construct encoding VP2 gene of infectious bursal disease virus and truncated HSP70 of Mycobacterium tuberculosis as a vaccine candidate was evaluated.
6. Development and evaluation of a Salmonella typhimurium flagellin based chimeric DNA vaccine against infectious bursal disease of poultry.

Subunit Vaccine

1. Recombinant epsilon toxoid of Clostridium perfringens as vaccine for enterotoxeimia.
2. Recombinant OMPs of Pasteurella multocida as vaccine candidate against HS.
3. Development of sub-viral particles of infectious bursal disease virus in yeast expression system as vaccine candidate against infectious bursal disease.
4. Antigenic validation of recombinant hemagglutinin-neuraminidase protein of Newcastle disease virus expressed in Saccharomyces cerevisiae evaluated.
5. Protective effects of recombinant glycoprotein D based prime boost approach against duck enteritis virus in mice model carried out.

Therapeutics

1. Replication-defective adenoviruses and lentiviruses encoding siRNA targeting rabies virus (RV) nucleoprotein (N), glycoprotein (G) and polymerase (L) genes have been constructed and evaluated for their anti-rabies effect in vitro in BHK-21 cells and in vivo in mice.
2. DNA constructs encoding apoptotic genes have been developed as canine mammary tumor therapeutics.

Peptide-nucleic acids (PNAs)

1. Newcastle disease virus specific PNAs were designed and evaluated for their ability to inhibit virus replication as well as their use in detection of viral RNA.

Viral gene construct

1. Molecular pathways for apoptosis caused by NDV and CPV-2 elucidated.
2. Oncolytic potential of NDV was established in vivo in Rous sarcoma virus (RSV) induced tumour.
3. Oncolytic potential and molecular pathway involved in apoptosis cased by VP3 gene of CAV and NS1 gene of CPV-2 was established in vitro in HeLa cells.

Vector construction/Development

1. RNA-replicon based self replicating mammalian expression vector (pAlpha) developed for enhanced heterologous gene expression and DNA immunization.
2. Developed B0HV-1 as viral expression vector and evaluated using expression of red fluorescent protein.
3. Reverse genetics system derived from a classical swine fever virus field isolate developed for genetic manipulation of infectious CSFV cDNA clone.
4. Rescue of Newcastle disease virus strain R2B with green fluorescent protein as a way towards development of NDV as viral vaccine vector.
5. Tumor promoter specific gene constructs designed and characterized for specific expression and apoptosis in canine cancer cells.

Diagnostics

Recombinant antigen based

1. Recombinant VP2 antigen based latex agglutination test for sero-diagnosis of Infectious bursal disease of poultry was developed.
2. Recombinant haemagglutinin antigen developed in prokaryotic system for detection of antibodies against Newcastle disease of poultry.
3. Recombinant fiber antigen developed in prokaryotic system for detection of antibodies against Hydropericardium syndrome of poultry.
4. Recombinant nucleoprotein antigen based single serum dilution ELISA was developed in yeast expression system for sero monitoring of Newcastle disease of poultry by screening a total of 1182 sera samples.
5. Recombinant B cell antigens of Mycobacterium avium paratuberculosis for detection of antibodies in paratuberculosis infected animals using ELISA based assay.
6. Recombinant multi-antigenic proteins of Mycobacterium avium paratuberculosis as diagnostic reagent.
7. Recombinant T cell antigen an alternative to johnin (PPD) for testing in M.a.paratuberculosis (Map) sensitized animals.

PCR based

1. A single tube reverse-transcription (RT) PCR based diagnostic test was developed for detection of Rabies in suspected brain samples.
2. RT-PCR (one step and two steps both) were developed for detection of IBD,bluetongue, Newcastle disease and classical swine fever viruses from clinical samples.
3. RT-PCR in combination with RFLP was optimized for characterization of NDV isolates in lentogenic, mesogenic and velogenic strains and its pathotyping directly from clinical samples.
4. PCR based detection of BHV-1 (IBR) from bovine serum.
5. A rapid, sensitive and user friendly visual LAMP based assay was developed for detection of IBR virus in bovine semen.
6. PCR based detection of Mycobacterium avium paratuberculosis in clinical samples.
7. PCR based amplification and detection of Clostridium perfringens epsilon toxin from clinical samples.
8. PCR based amplification of Clostridium perfringens DNA for toxinogenic typing.
9. LAMP assays developed for rapid, sensitive and visual detection of BoHV-1 and Brucella in semen.
10. Real-time PCR based diagnostic test developed for detection of BoHV-1 and Brucella in semen.
11. Real-time LAMP assay developed for quantitative and qualitative estimation of avian reovirus.

Synthetic peptide based

1. PPR specific peptides identified for detection and quantification of PPRV specific antibodies
2. A lateral flow penside assay for PPR virus detection using high affinity monoclonal antibodies fabricated on colloidal gold particles was developed.
3. PNA based point-of-care visual test for viral RNA detection, quantification and pathotyping of NDV.

Biosensor based diagnostics

1. SPR based biosensor assays for label free, real time detection of serum cancer biomarkers in canine cancers developed.
2. SPR based biosensor assays for label free, real time detection of Brucella specific antibodies developed

Molecular characterization of pathogens

1. Classical swine fever virus lapinized vaccine strain and Indian field isolate (CSFV/IVRI/VB-131) have been characterized by complete genome sequencing.
2. BoHV-1 indian isolate number 216 has been characterized by complete genome sequencing.
3. Newcastle disease virus mesogenic strain R2B, lentogenic strain F and peacock strain 12 have been characterized by complete genome sequencing.
4. Peste des petits ruminants virus (Sungri/96 vaccine strain) has been characterized by complete genome sequencing.

Other Priority Areas - Stem Cell Biology

1. Established five mesenchymal stem cell lines of porcine origin and identified mesenchymal stem cells as resource cells for screening of ELOVL6 and SCD1 genes.
2. Generated putative iPS (induced pluripotent stem cells) of porcine origin.

Technology/patent developed

Patents Granted

1. Recombinant antigen based rapid sero-diagnosis of infectious bursal disease (IBD) – Dr. Sohini Dey, Dr. C. Madhan Mohan and Dr. J.M. Kataria (Patent No. 277404 ).
2. Synthetic peptide antigen for diagnosis of Peste des petits ruminants (PPR) - Dr Satish Kumar, Sameer Shrivastava, R.K.Singh, and M.P.Yadav (Patent No278655).
3. Multiple antigenic peptide (MAP) assay for detection of Peste des petits ruminants (PPR) virus specific antibodies - Dr Satish Kumar, Sameer Shrivastava, R.K.Singh, and M.P.Yadav (Patent No 278622).
4. “An artificially synthesized peptide”. Dr Satish Kumar, Dr. M.V.Bais, Dr. R.S.Kataria and Dr. M.P.Yadav. International DCT publication No. WO/2008/041047, US-patent application no. 12/311554 (8216843 B2)

Patents Filed

1. Pestivirus replicase based self-replicating RNA-replicon vector for heterologous gene expression in mammalian cells. Dr. Praveen K. Gupta and Dr. Chhabi Lal Patel (Patent filed vide No 3805/Del/2011).
2. A rapid sensitive and user friendly visual LAMP-based assay for detection of infectious bovine rhinotrachaitis (IBR) virus in bovine semen. Dr. Praveen K. Gupta, Dr. Sachin S. Pawar, Chetan D. Meshram, Dr. B.P. Mishra (Patent filed vide No 627/Del/2012).
3. Recombinant antigen based sero-diagnosis of Newcastle disease - Dr. C. Madhan Mohan, Dr. Sohini Dey and Dr. J.M. Kataria (Patent filed vide No. 628/DEL/2012).
4. Generation of Newcastle disease virus strain R2B from cDNAs by reverse genetics technology. Madhan Mohan, C. and Sohini Dey (Patent application No.300/DEL/2015 ).

Technologies developed

• Development of single serum dilution ELISA kit for sero diagnosis of Newcastle disease of chickens.
• Development of single serum dilution ELISA kit for sero diagnosis of Infectious bursal disease of chickens.
• Development of sub-viral particles of infectious bursal disease virus in yeast as a vaccine candidate.

Awards/Recognition in the last 5 years