| dc.contributor.author | Kumar, C.S. |
| dc.contributor.author | Sireesha, K. |
| dc.contributor.author | Rao, G.V. |
| dc.contributor.author | Reddy, A.S. |
| dc.contributor.author | Rambabu, C. |
| dc.contributor.author | Kumar, P.L. |
| dc.date.accessioned | 2019-12-04T11:14:36Z |
| dc.date.available | 2019-12-04T11:14:36Z |
| dc.date.issued | 2008 |
| dc.identifier.citation | Kumar, C.S., Sireesha, K., Rao, G.V., Reddy, A.S., Rambabu, C. & Kumar, P.L. (2008). Development and evaluation of immunochemical tools for diagnosis and quality control of Helicoverpa armigera nucleopolyhedrovirus (HaNPV). Indian Journal of Virology, 19(1), 108-109. |
| dc.identifier.issn | 0970-2822 |
| dc.identifier.uri | https://hdl.handle.net/20.500.12478/2808 |
| dc.description.abstract | Helicoverpa armigera nucleopolyhedrovirus (HaNPV,
Family: Baculoviridae; Genus: Nucleopolyhedrovirus) is a
natural pathogen of Helicoverpa armigera (Insecta:
Arthropoda) larvae and has proven to be a good candidate
for the biocontrol of this pest on legume crops. In this study
various immunochemical tools were developed using the
polyclonal antibodies raised against thepolyocclusion body
(POB) protein (polyhedrin) and evaluated for the detection
and quantification of HaNPV in insect larvae and viral
insecticide prepartions. Indirect immunofluorescence assay
and Western immunoblot assay were developed for detection
of POBs in homogeneates of HaNPV-infected larvae. Direct
antigen coating (DAC)-enzyme-linkedimmunosorbent assay
(ELISA) and Indirect Competitive (1C)-ELISA were
developed for detection and quantification of polyhedrin
protein in insect extracts. The sensitivity of DAC-ELISA
is 30 nglml of HaNPV polyhedrin in 5 pglml of insect total
protein extracts. But in DAC-ELISA there was competition
between insect and viral proteins for binding to the ELISA
plate surface reducing the sensitivity ofthe assay. To eliminate
this, IC-ELISA was developed, which has sensitivity of
0.156pglml of HaNPV pol yhedrin in 20 pglml of total insect
proteinkxtracts. The cbncentration of pdlyhedrin for 50%
competitive inhibition (IC5d was calculated to be 1.14 pgl
ml. This test is equally effective in detecting polyhedrins
of heterologous NPVs such as, Spodoptera litura
Nucleopolyhedrovirus (0.3 1 pglml) and Amsacta albistriga
Nucleopolyhedrovirus (0.32 pglml). A simple purification
protocol was standardized for extraction of total polyhedrin
from NPV preparations of 6x10~to 4.68x107 POBS Iml.
The purity of the extracted polyhedrin was assayed in SDSPAGE
and evaluated in both DAC as well as IC-ELISA with
sensitivity of 9.375x107 POBS Iml. The ELISA results were
comparable to light microscope counting of POBs.
Application of ELISA and Western immunoblot assay in
bioassay experiments suggested that the 4th instar larvae
is better for virus innoculation, and virus harvesting 9 days
after inoculation for maxumum virus yield, and less bacterial
contamination. These diagnostic tools are convenient, rapid
and inexpensive for routine detection and quantification of HaNPV. |
| dc.language.iso | en |
| dc.subject | Nucleopolyhedrovirus |
| dc.subject | Helicoverpa Armigera |
| dc.subject | Polyhedrin Protein |
| dc.subject | Dac-Elisa |
| dc.subject | Polyhedrin |
| dc.title | Development and evaluation of immunochemical tools for diagnosis and quality control of Helicoverpa armigera nucleopolyhedrovirus (HaNPV) |
| dc.type | Journal Article |
| dc.description.version | Peer Review |
| cg.contributor.affiliation | International Crops Research Institute for the Semi-Arid Tropics |
| cg.contributor.affiliation | All India Networking Project on Betel vine |
| cg.contributor.affiliation | Acharya Nagarjuna University |
| cg.contributor.affiliation | International Institute of Tropical Agriculture |
| cg.coverage.region | Asia |
| cg.coverage.region | Africa |
| cg.coverage.region | East Asia |
| cg.coverage.region | West Africa |
| cg.coverage.country | India |
| cg.coverage.country | Nigeria |
| cg.isijournal | ISI Journal |
| cg.authorship.types | CGIAR and developing country institute |
| cg.iitasubject | Pests Of Plants |
| cg.iitasubject | Plant Production |
| cg.iitasubject | Plant Diseases |
| cg.iitasubject | Disease Control |
| cg.iitasubject | Farm Management |
| cg.iitasubject | Food Security |
| cg.accessibilitystatus | Limited Access |
| local.dspaceid | 93909 |
