Characterization of immunosensor for early detection of Cucumber Mosaic Virus (CMV) detection in chili
Cucumber Mosaic Virus (CMV) is one of the major constraints towards cucurbit crops such as cucumbers, zucchinis, pumpkins, papayas and watermelons production, while for non-cucurbit crops such as chilies, tomatoes, spinach, lettuces, celeries, beans, tobaccos and weeds production in South and Sou...
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| Format: | Thesis |
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| Language: | English |
| Subjects: | |
| Online Access: | http://dspace.unimap.edu.my:80/xmlui/bitstream/123456789/76626/1/Page%201-24.pdf http://dspace.unimap.edu.my:80/xmlui/bitstream/123456789/76626/2/Full%20text.pdf http://dspace.unimap.edu.my:80/xmlui/bitstream/123456789/76626/3/Declaration%20Form.pdf |
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| Summary: | Cucumber Mosaic Virus (CMV) is one of the major constraints towards cucurbit crops
such as cucumbers, zucchinis, pumpkins, papayas and watermelons production, while
for non-cucurbit crops such as chilies, tomatoes, spinach, lettuces, celeries, beans,
tobaccos and weeds production in South and Southeast Asia. Based on previous studies
approximately 5-10 % annual losses of chili yield in Asia was caused by CMV, which
accounts for nearly RM 6.05 billion annual loss in chili production worldwide. This loss
has given a huge impact especially to the farmers if there are no serious actions taken.
Most farmers are using common approach which is visual observation to detect the
CMV on their crops despite the visual observation is difficult to identify a symptom
caused by the CMV since the visual of symptom depends on the concentration of virus
itself and because of that reason, this method is not reliable to eradicate this virus from
scratch. Therefore, a portable electrochemical immunosensor based cucumber mosaic
virus detection like screen-printed carbon electrode was developed and it can be
employed whether in laboratory and field that is essential. In addition, study on the
CMV disease and antibody activity was demonstrated to making sensor actively
recognize only the CMV molecules by using specify antibody. The CMV purification is
used to eliminate the impurities and then, optimization of the purified CMV was
performed using sandwich immunoassay format. The purification of antibody was
demonstrated to eliminate salt and other proteins and the purified antibody was
optimized using sandwich immunoassay format and direct immunoassay format. The
initial results showed the both purified substance possess high binding strength.
Subsequently, a purified antibody was conjugated with gold nanoparticles and the
conjugated solution was used for the immunosensor surface modification to change the
immunosensor surface properties. The electrical signal produced from the sensor
validation process was measured using chronoamperometric (CM) technique. By using
the same technique, the set voltage potential was spotted at 0.2 V and the LOD of
immunosensor was at 0.1 mg mL-1. After that, the immunosensor was tested with other
pathogens to verify the immunosensor selectivity. The initial study shows the
immunosensor fully identifies the purified CMV and did not react to other purified
pathogens whereby gives the lowest cross-reactivity. In the CMV screening the same
cross-reactivity method was executed and the crude chili leaves that taken around
MARDI plantation were used to detect CMV in the chili trees. The results of the CMV
screening show the presence of CMV in some samples. Thus, this research provides a
sensitive and selective detection tool to the farmers that allow an early detection on their
chili plantations. |
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