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231225s2018 xx |||||o 00| ||eng c |
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|a 10.1094/PDIS-08-17-1160-RE
|2 doi
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|a pubmed25n0976.xml
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|a (NLM)30673493
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|a DE-627
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|c DE-627
|e rakwb
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|a eng
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| 100 |
1 |
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|a Buonicontro, Dalila Sêni
|e verfasserin
|4 aut
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| 245 |
1 |
2 |
|a A Rapid Diagnostic for Detection of Aphelenchoides besseyi and A. fujianensis Based on Real-Time PCR
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|c 2018
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| 336 |
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|a Text
|b txt
|2 rdacontent
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| 337 |
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|a ƒaComputermedien
|b c
|2 rdamedia
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| 338 |
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|a ƒa Online-Ressource
|b cr
|2 rdacarrier
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| 500 |
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|a Date Completed 14.02.2019
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|a Date Revised 10.12.2019
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|a published: Print-Electronic
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|a Citation Status MEDLINE
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|a Aphelenchoides besseyi and A. fujianensis have been frequently found in mixed populations associated with forage grass seed in Brazil. The morphological similarity between both species has previously led A. fujianensis to be erroneously identified as A. besseyi. A. besseyi is a quarantine pest in many countries that import Brazilian forage seed; however, there is no current evidence suggesting that A. fujianensis is a plant-parasitic species. Two real-time polymerase chain reaction (qPCR) diagnostics were developed to detect each species and an operational envelope was established. A set of primers and hydrolysis probes for each species was designed targeting the large subunit (LSU) region. To assess their specificity, primers and probes sets were tested with samples of nontarget Aphelenchoides and Paraphelenchus sp. also frequently associated with forage seed. Experiments using dilutions of purified plasmid standards underpinned the sensitivity of the qPCR assays, which detected as few as 10 copies of target nematode ribosomal DNA. Thus, the developed diagnostics were sufficiently sensitive to detect DNA extracted from a fragment of a single target nematode. There was a positive correlation between copy number of the target species and nematode abundance, suggesting the potential of this method for quantification. Evidence of intra-individual variability among cloned sequences of the LSU region in a single A. besseyi population is also reported
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|a Journal Article
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4 |
|a Research Support, Non-U.S. Gov't
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4 |
|a Validation Study
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| 650 |
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7 |
|a DNA Primers
|2 NLM
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| 650 |
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|a DNA, Helminth
|2 NLM
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| 650 |
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7 |
|a DNA, Ribosomal
|2 NLM
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| 700 |
1 |
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|a Roberts, David Mark
|e verfasserin
|4 aut
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| 700 |
1 |
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|a Oliveira, Claudio Marcelo Gonçalves
|e verfasserin
|4 aut
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| 700 |
1 |
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|a Blok, Vivian Carol
|e verfasserin
|4 aut
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| 700 |
1 |
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|a Neilson, Roy
|e verfasserin
|4 aut
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| 700 |
1 |
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|a Oliveira, Rosângela D'arc De Lima
|e verfasserin
|4 aut
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| 773 |
0 |
8 |
|i Enthalten in
|t Plant disease
|d 1997
|g 102(2018), 3 vom: 01. März, Seite 519-526
|w (DE-627)NLM098181742
|x 0191-2917
|7 nnns
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| 773 |
1 |
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|g volume:102
|g year:2018
|g number:3
|g day:01
|g month:03
|g pages:519-526
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| 856 |
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|u http://dx.doi.org/10.1094/PDIS-08-17-1160-RE
|3 Volltext
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