Details
| Original language | English |
|---|---|
| Pages (from-to) | 548-552 |
| Number of pages | 5 |
| Journal | Zeitschrift fur Pflanzenkrankheiten und Pflanzenschutz |
| Volume | 107 |
| Issue number | 5 |
| Publication status | Published - 2000 |
Abstract
A reverse transcription-polymerase chain reaction-enzyme-linked immunosorbent assay (RT-PCR-ELISA) for the detection of Chrysanthemum stunt viroid (CSVd) in Argyranthemum frutescens was developed. A sequence-specific, modified primer pair (5'-end labelled with digoxigenin or biotin) was used, defining a target sequence of 281 bp. The digoxigenin and biotin-labeled PCR products were directly bound to streptavidin-coated microtiter plates and detected using anti-dixoxigenin Fab fragments conjugated with alkaline phosphatase. The described technique does not require denaturation and hybridization of a detection probe to the amplified products and operates without the use of hazardous organic solvents and toxic or mutagenic stains. Only 6.5 h are required for the detection of CSVd from infected tissue. This ELISA technique could potentially replace common electrophoretic techniques for the detection of PCR products.
Keywords
- Argyranthemum, csvd, Detection, RT-PCR-ELISA
ASJC Scopus subject areas
- Agricultural and Biological Sciences(all)
- Plant Science
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In: Zeitschrift fur Pflanzenkrankheiten und Pflanzenschutz, Vol. 107, No. 5, 2000, p. 548-552.
Research output: Contribution to journal › Article › Research › peer review
}
TY - JOUR
T1 - Detection of Chrysanthemum stunt viroid (CSVd) in cultivars of Argyranthemum frutescens by RT-PCR-ELISA
AU - Menzel, W.
AU - Maiss, E.
PY - 2000
Y1 - 2000
N2 - A reverse transcription-polymerase chain reaction-enzyme-linked immunosorbent assay (RT-PCR-ELISA) for the detection of Chrysanthemum stunt viroid (CSVd) in Argyranthemum frutescens was developed. A sequence-specific, modified primer pair (5'-end labelled with digoxigenin or biotin) was used, defining a target sequence of 281 bp. The digoxigenin and biotin-labeled PCR products were directly bound to streptavidin-coated microtiter plates and detected using anti-dixoxigenin Fab fragments conjugated with alkaline phosphatase. The described technique does not require denaturation and hybridization of a detection probe to the amplified products and operates without the use of hazardous organic solvents and toxic or mutagenic stains. Only 6.5 h are required for the detection of CSVd from infected tissue. This ELISA technique could potentially replace common electrophoretic techniques for the detection of PCR products.
AB - A reverse transcription-polymerase chain reaction-enzyme-linked immunosorbent assay (RT-PCR-ELISA) for the detection of Chrysanthemum stunt viroid (CSVd) in Argyranthemum frutescens was developed. A sequence-specific, modified primer pair (5'-end labelled with digoxigenin or biotin) was used, defining a target sequence of 281 bp. The digoxigenin and biotin-labeled PCR products were directly bound to streptavidin-coated microtiter plates and detected using anti-dixoxigenin Fab fragments conjugated with alkaline phosphatase. The described technique does not require denaturation and hybridization of a detection probe to the amplified products and operates without the use of hazardous organic solvents and toxic or mutagenic stains. Only 6.5 h are required for the detection of CSVd from infected tissue. This ELISA technique could potentially replace common electrophoretic techniques for the detection of PCR products.
KW - Argyranthemum
KW - csvd
KW - Detection
KW - RT-PCR-ELISA
UR - http://www.scopus.com/inward/record.url?scp=0033756896&partnerID=8YFLogxK
M3 - Article
AN - SCOPUS:0033756896
VL - 107
SP - 548
EP - 552
JO - Zeitschrift fur Pflanzenkrankheiten und Pflanzenschutz
JF - Zeitschrift fur Pflanzenkrankheiten und Pflanzenschutz
SN - 0340-8159
IS - 5
ER -