2007
DOI: 10.1080/07060660709507456
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Polymerase chain reaction detection of greening bacterium (Candidatus Liberibacter asiaticus) and Citrus mosaic virus in citrus tissues, by means of a simplified template-preparation protocol

Abstract: Application of a simplified protocol for nucleic acid preparation for polymerase chain reaction (PCR) detection of greening bacterium (Candidatus Liberibacter asiaticus; Cla) and Citrus mosaic virus (CMBV) associated with citrus is described. Crude extracts of citrus tissues in NaOH -ethylenediaminetetraacetic acid solution, prepared without the use of liquid nitrogen, were spotted on a nitrocellulose membrane (NCM) and then eluted in water at 80°C for 10 min. Both the greening bacterium and CMBV were detected… Show more

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Cited by 6 publications
(4 citation statements)
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“…In some herbaceous plants crude extract of denaturized sap is also used for detection purpose but as citrus is a woody plant its crude extract may contains PCR inhibitors and therefore direct plant sap may not be used for detection purpose. There was a report of successful detection of citrus pathogens individually using membrane based technique (Baranwal et al, 2007). In the present study however, we have developed a duplex PCR for simultaneous detection of CMBV and 'Ca.…”
Section: Shelf-life Evaluation Of Spotted Ncm Discmentioning
confidence: 82%
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“…In some herbaceous plants crude extract of denaturized sap is also used for detection purpose but as citrus is a woody plant its crude extract may contains PCR inhibitors and therefore direct plant sap may not be used for detection purpose. There was a report of successful detection of citrus pathogens individually using membrane based technique (Baranwal et al, 2007). In the present study however, we have developed a duplex PCR for simultaneous detection of CMBV and 'Ca.…”
Section: Shelf-life Evaluation Of Spotted Ncm Discmentioning
confidence: 82%
“…However, most of the PCR assays developed for CMBV and 'Ca. Las', involves either multistep DNA isolation protocol or expensive commercial kit (La Notte et al, 1997;Baranwal et al, 2007). It is also not practical to rapidly index large scale field or nursery samples for pathogen testing.…”
Section: Research Articlementioning
confidence: 99%
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“…The evaluation was also done using Taq and Klen Taq enzymes and Klen Taq as was expected gave better intensity of the bands obtained in 1% agarose gel. Similar evaluation for primer has also been used by other researchers with different viruses (Singh and Nie, 2003;Harakava et al, 2000;Das, 2004;Manjunath et al, 2008;Baranwal et al, 2007;Khairulmazmi et al, 2008;Naoya et al, 2008;Thiara et al, 2009;Albrecht and Kim Bowman, 2009;Deng et al, 2012).…”
Section: Determination Of Quality Of Dna Isolated By Three Methodsmentioning
confidence: 88%