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231225s2020 xx |||||o 00| ||eng c |
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|a 10.1094/PDIS-05-19-0931-RE
|2 doi
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|a pubmed24n1013.xml
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|a (NLM)31790641
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|a DE-627
|b ger
|c DE-627
|e rakwb
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|a eng
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| 100 |
1 |
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|a Bao, Minli
|e verfasserin
|4 aut
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| 245 |
1 |
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|a Enhancing PCR Capacity To Detect 'Candidatus Liberibacter asiaticus' Utilizing Whole Genome Sequence Information
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|c 2020
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|a Text
|b txt
|2 rdacontent
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|a ƒaComputermedien
|b c
|2 rdamedia
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| 338 |
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|a ƒa Online-Ressource
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|2 rdacarrier
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|a Date Completed 21.02.2020
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|a Date Revised 21.02.2020
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|a published: Print-Electronic
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|a Citation Status MEDLINE
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|a 'Candidatus Liberibacter asiaticus' (CLas) is an unculturable α-proteobacterium associated with citrus Huanglongbing (HLB; yellow shoot disease). PCR procedures that accurately confirm or exclude CLas infection in citrus tissue/Asian citrus psyllid (ACP) samples are critical for HLB management. When CLas was described in 1994, a 23-bp signature oligonucleotide sequence (OI1) in the 16S rRNA gene (rrs, three genomic copies) was identified based on Sanger sequencing. OI1 contains single nucleotide polymorphisms (SNPs) distinguishing CLas from non-CLas species. The SNPs were used to design the primer HLBas, a key primer for a commonly used TaqMan PCR system (HLBas-PCR) for CLas detection. Recent developments in next-generation sequencing technology have led to the identification of 15 CLas whole genome sequence strains (WGSs). Analyses of CLas WGSs have generated a significant amount of biological information that could help to improve CLas detection. Utilizing the WGS information, this study re-evaluated the sequence integrity of OI1/HLBas and identified and/or confirmed a missing nucleotide G in the two primers. Replacement primers for OI1 and HLBas are proposed. At low cycle threshold (Ct) values (e.g., <30), HLBas-PCR remained reliable in CLas determination. At high Ct values (e.g., >30), HLBas-PCR alone was unreliable in differentiating whether samples contain low CLas titers or whether CLas is not present. The availability of ribonucleotide reductase (RNR)-PCR derived from the five-copy nrdB gene helped to resolve this problem. To further enhance low CLas titer detection, a 4CP-PCR system, based on a four-copy genomic locus, was developed. Evaluation of 107 HLB samples (94 citrus and 13 ACP) showed that 4CP-PCR was more sensitive than HLBas-PCR and shared similar sensitivity with RNR-PCR
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|a Journal Article
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|a pathogen detection
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|a prokaryotes
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4 |
|a techniques
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| 650 |
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|a RNA, Ribosomal, 16S
|2 NLM
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| 700 |
1 |
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|a Zheng, Zheng
|e verfasserin
|4 aut
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| 700 |
1 |
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|a Sun, Xiaoan
|e verfasserin
|4 aut
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| 700 |
1 |
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|a Chen, Jianchi
|e verfasserin
|4 aut
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| 700 |
1 |
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|a Deng, Xiaoling
|e verfasserin
|4 aut
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| 773 |
0 |
8 |
|i Enthalten in
|t Plant disease
|d 1997
|g 104(2020), 2 vom: 08. Feb., Seite 527-532
|w (DE-627)NLM098181742
|x 0191-2917
|7 nnns
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| 773 |
1 |
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|g volume:104
|g year:2020
|g number:2
|g day:08
|g month:02
|g pages:527-532
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| 856 |
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|u http://dx.doi.org/10.1094/PDIS-05-19-0931-RE
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