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Use of the reverse transcription-polymerase chain reaction for the detection of pelargonium flower break carmovirus
Year:
1997
Source of publication :
Journal of Phytopathology
Authors :
Franck, Andre
;
.
Gera, Abdullah
;
.
Loebenstein, Gad
;
.
Volume :
145
Co-Authors:
Franck, A., Department of Virology, Agricultural Research Organization, Volcani Center, Bet Dagan 50250, Israel
Loebenstein, G., Department of Virology, Agricultural Research Organization, Volcani Center, Bet Dagan 50250, Israel
Gera, A., Department of Virology, Agricultural Research Organization, Volcani Center, Bet Dagan 50250, Israel
Facilitators :
From page:
235
To page:
238
(
Total pages:
4
)
Abstract:
A polymerase chain reaction (PCR)-based assay was used to detect pelargonium flower break carmovirus (PFBV) in total RNA extractions made from infected Pelargonium plants. Extracts were reverse transcribed (RT) and the resultant cDNA was amplified by PCR, using oligonucleotide primers specific for 343, 510 and 832 base pair fragments of the RNA-dependent RNA polymerase gene of PFBV. The specificity and sensitivity of RT-PCR were compared with the enzyme-linked immunosorbent assay (ELISA) for the detection of PFBV in Pelargonium tissues. The virus could be detected efficiently in high dilutions of sap from infected plants and at low concentrations of purified virus. Although ELISA is a powerful tool for virus detection, RT-PCR was over 1000 times more sensitive in detecting PFBV in leaf extracts of infected Pelargonium than was ELISA. The limit of detecting PFBV RNA by RT-PCR was 200 fg, compared with 200 pg of virus by ELISA.
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DOI :
Article number:
Affiliations:
Database:
Scopus
Publication Type:
article
;
.
Language:
English
Editors' remarks:
ID:
30649
Last updated date:
02/03/2022 17:27
Creation date:
17/04/2018 00:56
Scientific Publication
Use of the reverse transcription-polymerase chain reaction for the detection of pelargonium flower break carmovirus
145
Franck, A., Department of Virology, Agricultural Research Organization, Volcani Center, Bet Dagan 50250, Israel
Loebenstein, G., Department of Virology, Agricultural Research Organization, Volcani Center, Bet Dagan 50250, Israel
Gera, A., Department of Virology, Agricultural Research Organization, Volcani Center, Bet Dagan 50250, Israel
Use of the reverse transcription-polymerase chain reaction for the detection of pelargonium flower break carmovirus
A polymerase chain reaction (PCR)-based assay was used to detect pelargonium flower break carmovirus (PFBV) in total RNA extractions made from infected Pelargonium plants. Extracts were reverse transcribed (RT) and the resultant cDNA was amplified by PCR, using oligonucleotide primers specific for 343, 510 and 832 base pair fragments of the RNA-dependent RNA polymerase gene of PFBV. The specificity and sensitivity of RT-PCR were compared with the enzyme-linked immunosorbent assay (ELISA) for the detection of PFBV in Pelargonium tissues. The virus could be detected efficiently in high dilutions of sap from infected plants and at low concentrations of purified virus. Although ELISA is a powerful tool for virus detection, RT-PCR was over 1000 times more sensitive in detecting PFBV in leaf extracts of infected Pelargonium than was ELISA. The limit of detecting PFBV RNA by RT-PCR was 200 fg, compared with 200 pg of virus by ELISA.
Scientific Publication
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