Detection of blueberry scorch virus strain NJ2 by reverse transcriptase-polymerase chain reaction amplification

A reverse transcription-polymerase chain reaction (RT-PCR) assay has been developed for the detection of strain NJ2 of blueberry scorch carlavirus (BBScV), which causes an economically important disease of blueberry. Four extraction methods were compared for their ability to yield nucleic acid from blueberry tissue that could be amplified by specific RT-PCR primers. Two extraction methods were found suitable when they were followed by an Elutip-r column purification step. These procedures were successful with blueberry blossoms and leaves. RT-PCR was shown to be considerably more sensitive than spot hybridization using a P-32-labeled DNA probe. These purification systems enable use of nucleic acid-based RNA virus detection from blueberry.