Production of monoclonal antibodies for detection and identification of strains of potato virus Y

Ellis, Peter; Stace-Smith, R.; Bowler, G.; MacKenzie, Donald J.

doi:10.1080/07060669609500657

Cited by 58 publications

(36 citation statements)

References 13 publications

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“…PVY C and PVY O are serologically distinguishable, and it is possible to distinguish these using monoclonal antibodies that recognise a specific epitopes [42,58]. However, despite the improved specificity associated with MAbs, these diagnostic tools have not resolved the problems associated with strain designation [15,58]. Molecular studies of the coat protein cistron on a range of PVY isolates has resolved the isolates into three groups [38] and confirmed that PVY isolates inducing PTNRD belong to the PVY N strain group [5].…”

Section: Introductionmentioning

confidence: 90%

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Characterisation of potato virus Y isolates from Iran

et al. 2010

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A survey of Potato virus Y (PVY) was conducted in cultivated fields in six Iranian provinces between January 2005 to July 2007. Two hundred samples from potato and tomato were collected and analyzed by enzyme-linked immunosorbent assay (ELISA) for potyviruses. Almost one fourth of the samples were found to be infected by PVY. Analysis of these PVY-positive samples using three monoclonal antibodies (MAbs) facilitating the simultaneous detection of three main strains namely the ordinary (PVY(O)), strain (PVY(N)) and C (PVY(C)) strains. However, the fourth strain (PVY(NTN)) and some others recombinant isolates were also identified by molecular methods. Host range and symptoms analysis using sap inoculation of four different strains of PVY onto a range of plants revealed that the four strains showed biological properties that seemed to be consistent with their molecular grouping. Fourteen isolates of PVY were chosen based on the host and geographical location, primer specificity and serology for further biological and molecular characterisation. The coat protein (CP) and P1 genes and 3'-non-translated region (3'NTR) from 14 representative isolates were sequenced and analysed with the sequences available in GenBank. Composite analysis of the P1, CP and 3'-UTR sequences with all full genome sequences of PVY revealed that there are three potential strains of PVY in Iran, PVY(O), PVY(N)-W and PVY(NTN). Isolate KER.SA(N) was the most divergent of all the 14 isolates reacted with PVY(N) specific MAbs but grouped with PVY(O) strains in maximum likelihood phylogentic analysis. The PVY(NTN) isolates from Iran more closely related to the European than North American PVY(NTN) isolates.

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Section: Introductionmentioning

confidence: 90%

“…The introduction of strain-specific MAbs represents a significant advancement in the detection and diagnosis of the various PVY strains [15,16,52]. PVY C and PVY O are serologically distinguishable, and it is possible to distinguish these using monoclonal antibodies that recognise a specific epitopes [42,58].…”

Section: Introductionmentioning

confidence: 99%

Characterisation of potato virus Y isolates from Iran

et al. 2010

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“…Even though the role of these amino acids in the determination of the PVY N/NTN serotype cannot be concluded, they can be used to identify this serotype. Monoclonal antibodies are not always able to discriminate PVY strains, due to the cross reaction of these antibodies and the presence of recombination events in the CP of some PVY isolates [14,15,42]. Thus, these amino acid markers can be used to distinguish the CP of PVY isolates and thereby their serotypes and phylogenetic relationships.…”

Section: Discussionmentioning

confidence: 97%

“…MAb2 is a monoclonal antibody, described by McDonald and Kristjansson [16], which is capable of identifying isolates of PVY O and PVY C without any cross reaction with isolates of PVY N [10,14]. Another monoclonal antibody, 1F5, reported by Ellis et al [14] could be used to identify all isolates of PVY N with only a few isolates of the common strain PVY O cross reacting.…”

Section: Introductionmentioning

confidence: 97%

“…However, using monoclonal antibodies, PVY isolates could be differentiated into two main serological groups: PVY O,C serotype, which includes PVY O , PVY C , PVY Z , and PVY N W (designated as PVY N:O in North America), and PVY N serotype, hereafter referred to as PVY N/NTN , which includes PVY N and PVY NTN [8][9][10][11][12][13]. A range of monoclonal antibodies have been reported to identify PVY serotypes [14][15][16][17]. MAb2 is a monoclonal antibody, described by McDonald and Kristjansson [16], which is capable of identifying isolates of PVY O and PVY C without any cross reaction with isolates of PVY N [10,14].…”

Section: Introductionmentioning

confidence: 99%

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A point mutation changes the serotype of a Potato virus Y isolate; genomic determination of the serotype of PVY strains

2007

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A Syrian isolate of Potato virus Y (PVY), named PVY-12, reacted to two monoclonal antibodies that are specific to PVY(O,C) and PVY(N) strains, although its coat protein (CP) belongs to the PVY(N) strain. Analysis of the CP of PVY-12 revealed that a point mutation in its N terminus switched it from PVY(N)-like to PVY(O)-like at this position. This mutation changed the second nucleotide of the codon that encodes the 29th amino acid of the CP of PVY-12 from A to G, which resulted in one amino acid substitution from Glu(29 )to Gly(29). The role of Gly(29) in the binding of PVY-12 to PVY(O,C)-specific monoclonal antibody was confirmed by gene expression in Escherichia coli. The N terminus of the CP gene of PVY-12 and another PVY isolate of the N serotype with identical CP to PVY-12 except for one amino acid substitution from Gly(29 )to Glu(29) was cloned and expressed in E. coli using a pUC18 vector. Resulting antigens showed similar reactivity to the relevant antibodies as same as the native CPs of these two isolates. Further analysis of the CP of PVY isolates showed that Gly(29) was conserved in the CP of PVY(O), PVY(C), PVY(N)W, and non-potato isolates of PVY while Gln(17) and Glu(31 )were conserved in the CP of PVY(N/NTN). Therefore, these amino acids are characteristic of the CP for these strain groups and subgroups in agreement with the serotype and phylogenetic relationships previously determined.

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Potato Viruses and their Management

Khurana

Diseases of Fruits and Vegetables: Volume II

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Production of monoclonal antibodies for detection and identification of strains of potato virus Y

Cited by 58 publications

References 13 publications

Characterisation of potato virus Y isolates from Iran

Characterisation of potato virus Y isolates from Iran

A point mutation changes the serotype of a Potato virus Y isolate; genomic determination of the serotype of PVY strains

Potato Viruses and their Management

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