Immunosensor-based detection of tungro disease in rice plant

Rice tungro disease is the major constraint, caused by a combination of rice tungro spherical virus (RTSV) and rice tungro bacilliform virus (RTBV). Major outbreaks of tungro have been occurred in countries of South and Southeast Asia, mediated by a viral carrier green leafhopper. To prevent serious...

Full description

Bibliographic Details
Main Author: Uda, Muhammad Nur Aiman
Format: Thesis
Language:English
Published: 2015
Subjects:
Online Access:http://psasir.upm.edu.my/id/eprint/56694/1/FK%202015%2049.pdf
_version_ 1825931466247241728
author Uda, Muhammad Nur Aiman
author_facet Uda, Muhammad Nur Aiman
author_sort Uda, Muhammad Nur Aiman
collection UPM
description Rice tungro disease is the major constraint, caused by a combination of rice tungro spherical virus (RTSV) and rice tungro bacilliform virus (RTBV). Major outbreaks of tungro have been occurred in countries of South and Southeast Asia, mediated by a viral carrier green leafhopper. To prevent serious outbreaks, detection of Tungro disease requires a fast, simple and sensitive method. The current study was initiated by transferring viruses from the green leafhopper to susceptible plant host varieties (Y1286 and MR81 for RTSV and RTBV, respectively). After inoculation process, both viruses were purified and spectrophotometrically measured as 1.363 and 1.6075 mg mL-1 for RTBV,1.227 and 1.6075 mg mL-1 for RTSV. Viral particles were observed by transmission electron microscopy, where RTBV was observed as 168 mm in length with a bacilliform, whereas RTSV appeared with 35 mm in diameter,showed rod-shaped rounded ends. Next, pure viruses were immunized in White New Zealand rabbits and antibody was analyzed on enzyme-linked immunoassay (ELISA) surface. Analysis with RTBV showed that the second bleed has the highest titer, whereas for RTSV bleed 1 has the highest titer (1.6960 mg mL-1 and 2.3251 mg mL-1). Analysis on screen-printed carbon electrode (SPCE) was incorporated with 0.075 M pyrolle monomer where electro-polymerization process occurred at 0.9 V amperometrically for 20 min. Chronoamperometry measurements showed the best potential to be used is located at 0.2 V, for both RTBV and RTSV. In addition, the antibody immobilized surfaces of SPCE were analyzed by scanning electron microscope. A linear standard curve for each virus obtained based on current measurement (μA), where R2 values were 0.9755 and 0.967, indicates a higher sensitivity of immunosensor developed. Cross reactivity studies, showed the specificity of the antibodies with a low cross-reactivity, although RTBV and RTSV it-self manifested strongest serological cross-reactivity.
first_indexed 2024-03-06T09:27:10Z
format Thesis
id upm.eprints-56694
institution Universiti Putra Malaysia
language English
last_indexed 2024-12-09T02:16:28Z
publishDate 2015
record_format dspace
spelling upm.eprints-566942024-10-22T01:03:42Z http://psasir.upm.edu.my/id/eprint/56694/ Immunosensor-based detection of tungro disease in rice plant Uda, Muhammad Nur Aiman Rice tungro disease is the major constraint, caused by a combination of rice tungro spherical virus (RTSV) and rice tungro bacilliform virus (RTBV). Major outbreaks of tungro have been occurred in countries of South and Southeast Asia, mediated by a viral carrier green leafhopper. To prevent serious outbreaks, detection of Tungro disease requires a fast, simple and sensitive method. The current study was initiated by transferring viruses from the green leafhopper to susceptible plant host varieties (Y1286 and MR81 for RTSV and RTBV, respectively). After inoculation process, both viruses were purified and spectrophotometrically measured as 1.363 and 1.6075 mg mL-1 for RTBV,1.227 and 1.6075 mg mL-1 for RTSV. Viral particles were observed by transmission electron microscopy, where RTBV was observed as 168 mm in length with a bacilliform, whereas RTSV appeared with 35 mm in diameter,showed rod-shaped rounded ends. Next, pure viruses were immunized in White New Zealand rabbits and antibody was analyzed on enzyme-linked immunoassay (ELISA) surface. Analysis with RTBV showed that the second bleed has the highest titer, whereas for RTSV bleed 1 has the highest titer (1.6960 mg mL-1 and 2.3251 mg mL-1). Analysis on screen-printed carbon electrode (SPCE) was incorporated with 0.075 M pyrolle monomer where electro-polymerization process occurred at 0.9 V amperometrically for 20 min. Chronoamperometry measurements showed the best potential to be used is located at 0.2 V, for both RTBV and RTSV. In addition, the antibody immobilized surfaces of SPCE were analyzed by scanning electron microscope. A linear standard curve for each virus obtained based on current measurement (μA), where R2 values were 0.9755 and 0.967, indicates a higher sensitivity of immunosensor developed. Cross reactivity studies, showed the specificity of the antibodies with a low cross-reactivity, although RTBV and RTSV it-self manifested strongest serological cross-reactivity. 2015-12 Thesis NonPeerReviewed text en http://psasir.upm.edu.my/id/eprint/56694/1/FK%202015%2049.pdf Uda, Muhammad Nur Aiman (2015) Immunosensor-based detection of tungro disease in rice plant. Masters thesis, Universiti Putra Malaysia. Rice tungro spherical virus Rice - Diseases and pests - Control Immunospecificity
spellingShingle Rice tungro spherical virus
Rice - Diseases and pests - Control
Immunospecificity
Uda, Muhammad Nur Aiman
Immunosensor-based detection of tungro disease in rice plant
title Immunosensor-based detection of tungro disease in rice plant
title_full Immunosensor-based detection of tungro disease in rice plant
title_fullStr Immunosensor-based detection of tungro disease in rice plant
title_full_unstemmed Immunosensor-based detection of tungro disease in rice plant
title_short Immunosensor-based detection of tungro disease in rice plant
title_sort immunosensor based detection of tungro disease in rice plant
topic Rice tungro spherical virus
Rice - Diseases and pests - Control
Immunospecificity
url http://psasir.upm.edu.my/id/eprint/56694/1/FK%202015%2049.pdf
work_keys_str_mv AT udamuhammadnuraiman immunosensorbaseddetectionoftungrodiseaseinriceplant