The Comparison of Antibodies Raised Against PLRV with Two Different Approaches - Viral Particles Purification and Recombinant Production of CP

Dg, Aseel; Ee, Hafez

doi:10.4172/2157-7471.1000407

Cited by 3 publications

(2 citation statements)

References 28 publications

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“…Many recombinant polyclonal antibodies specific to plant viruses have been previously generated for serological detection of viruses such as Tomato spotted wilt virus (Vaira et al 1996), Potato virus Y (Folwarczna et al 2008), Alfalfa mosaic virus (Khatabi et al 2012), and Wheat streak mosaic virus (Tatineni et al 2014). Recombinant antisera for Egyptian isolates of both Potato virus X (Soliman et al 2006) and PLRV (Aseel and Hafez 2017;El-Attar et al 2010) have also been produced. In our experiment, E. coli was used to express His.MBP-PLRV-MP from which PLRV-MP fusion protein was purified and anti-MP PLRV successfully produced.…”

Section: Discussionmentioning

confidence: 99%

“…2 Titer determination of PLRV-MP antiserum by Western blotting. Antiserum used at 10 different dilutions (1:4000, 1:8000, 1:10000, 1:16000, 1:20000, 1:32000, 1:40000, 1:64000, 1:80000, and 1:128000) against PLRV, each of which consists of a left lane of negative control and a right lane containing proteins extracted from PLRV-infected Nicotiana benthamiana leaves most important techniques for virus detection (Aseel and Hafez 2017) and the antiserum (either monoclonal or polyclonal) is the basis for the serology test. In most cases, the anti-coat protein (CP) antisera are used for virus detection; however, virus detection by the anti-CP antisera may not show whether the virus is in active (replication) or inactive stage.…”

Section: Discussionmentioning

confidence: 99%

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Development of polyclonal antiserum against movement protein from Potato leafroll virus and its application for the virus detection

et al. 2019

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The serological method is one of the most important techniques extensively used in crop production to detect different pathogens, especially plant viruses. An antiserum is essential for serological tests. The 17 kDa movement protein (MP) of Potato leafroll virus (PLRV) is related to the membranous structures and localized to the plasmodesmata, but there is no report on preparation of PLRV-MP antiserum for detection of PLRV. To prepare PLRV-MP antiserum, reverse transcription polymerase chain reaction (RT-PCR) was carried out to amplify the PLRV-MP gene, which was constructed into a prokaryotic vector to express the protein in Escherichia coli for immunization of rabbits, after purification. Western blotting revealed that this developed antiserum could effectively detect PLRV, but with better results from the perspective of color development and economics by using antiserum at the ratio range of 1:10000 to 1:40000, presenting high sensitivity and specificity to PLRV. The serological detection results for PLRV of the field samples were identical to the RT-PCR detection. This is the first report on the development of PLRV-MP antiserum that has been successfully used for both laboratory and field detection of PLRV. The results provide a fundamental tool for further research on the function of PLRV-MP.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Development of polyclonal antiserum against movement protein from Potato leafroll virus and its application for the virus detection

et al. 2019

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Heterologous expression of pepper mild mottle virus coat protein encoding region and its application in immuno-diagnostics

2020

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Detection and identification of a new isolate of Grapevine fanleaf Virus naturally infecting Grapevine plants in Egypt using qReal Time-PCR

Aseel

El-Aziz

Hafez

2019

Novel Research in Microbiology Journal

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Grapevine fanleaf virus (GFLV) is a member of the genus Nepovirus in the family Comoviridae, a widely distributed virus responsible for grapevine (Vitis vinifera) degeneration. This virus causes serious economic losses by reducing grape crop yield. The Quantitative Real-Time Reverse Transcription-Polymerase Chain Reaction (qReal Time-PCR) assay was carried out on (GFLV) recovered from infected grapevines leaves at Alexandria, Egypt. A 606 bp fragment of the GFLV RNA-2 coat protein (CP) gene was amplified and then sequenced. Results of reactions of diagnostic hosts were observed on Gomphrena globosa, which developed systemic mottling, leaves twisting and necrotic spots during spring, whereas Chenopodium amaranticolor induced systemic mottling and leaf deformation, and its sap seemed relatively insensitive to the inhibitors of infection. Mottling of Glycine max was detected after inoculation, but inoculation of Nicotiana glutinosa didn't induce any symptoms. This study aimed to detect and identify a new isolate of GFLV-DA3 from Egypt using biological and molecular tools.

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The Comparison of Antibodies Raised Against PLRV with Two Different Approaches - Viral Particles Purification and Recombinant Production of CP

Cited by 3 publications

References 28 publications

Development of polyclonal antiserum against movement protein from Potato leafroll virus and its application for the virus detection

Development of polyclonal antiserum against movement protein from Potato leafroll virus and its application for the virus detection

Heterologous expression of pepper mild mottle virus coat protein encoding region and its application in immuno-diagnostics

Detection and identification of a new isolate of Grapevine fanleaf Virus naturally infecting Grapevine plants in Egypt using qReal Time-PCR

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