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Use of the Reverse Transcription‐Polymerase Chain Reaction for the Detection of Pelargonium Flower Break Carmovirus
Author(s) -
Franck A.,
Loebenstein G.,
Gera A.
Publication year - 1997
Publication title -
journal of phytopathology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.53
H-Index - 60
eISSN - 1439-0434
pISSN - 0931-1785
DOI - 10.1111/j.1439-0434.1997.tb00392.x
Subject(s) - biology , polymerase chain reaction , microbiology and biotechnology , reverse transcriptase , reverse transcription polymerase chain reaction , virus , rna , real time polymerase chain reaction , virology , complementary dna , oligonucleotide , serial dilution , gene , polymerase , rna extraction , messenger rna , genetics , medicine , alternative medicine , pathology
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.