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PURIFICATION AND ISOLATION OF PAPAYA RING SPOT VIRUS AND IT?S IMMUNOLOGICAL AND MOLECULAR DETECTION

KrishiKosh

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Title PURIFICATION AND ISOLATION OF PAPAYA RING SPOT VIRUS AND IT?S IMMUNOLOGICAL AND MOLECULAR DETECTION
 
Creator MUSKE DEEPA, NAVANATH
 
Contributor ANITHA, PETER
 
Description Papaya (Carica papaya) is one of the major horticultural crops of India. In recent years, the production of papaya is greatly affected by a disease, caused by Papaya Ring Spot Virus (PRSV). It has an extensive host range infecting different crops and weeds. In the present study, the PRSV virus was purified and confirmed by electron microscopy, the dimensions of virus was 700-720 nm long, with a diameter of 12-13 nm.
Polyclonal antibodies were produced against PRSV in rabbits and a direct and indirect plate Enzyme Linked Immunosorbent Assay (ELISA) and Dot- ELISA standardized for virus detection.The antigen as a purified virus was used at dilutions of 1:10, 1:20, 1:50, primary antibody at 1:500, 1:1000, 1:2000, 1:4000, 1:8000, 1:16000 and secondary antibody (commercially available antirabbit immunoglobulin IgG conjugated with alkaline phosphatase) at 1:1000 and 1:2000 dilution, control included sample from healthy plants. The optimum OD reading at 405 nm for direct plate ELISA was 1.032 for 1:20 and 1.134 for 1:10 and 1.338 and 1.016 in 1:50, antigen dilution for indirect plate ELISA, at1:1000 and 1:2000 secondary antibody dilution respectively and 1:16000 dilution of primary antibody. In direct and indirect DOT-ELISA the titer was 1:50 of antigen, 1:16000 of primary antibody and 1:2000 of secondary antibody. No colour was seen in control. Reverse Transcriptase-Polymerase Chain Reaction (RT-PCR) amplification of PRSV Coat Protein (CP) and Helper Component-Pro (HC-Pro) gene using specific primers resulted in partial amplification of 534bp and 700bp fragments of the respective genes.
 
Date 2016-11-29T13:45:17Z
2016-11-29T13:45:17Z
2012-08-12
 
Type Thesis
 
Identifier Th-10333
http://krishikosh.egranth.ac.in/handle/1/88136
 
Language en
 
Format application/pdf
 
Publisher University of Agricultural Sciences GKVK, Bangalore