Issue link: http://read.dmtmag.com/i/100792
The Benefits of Real-Time PCR Technology delivers sensitive detection of grapevine viruses BY ANNA-LIISA FABRITIUS, PH.D., AND LANA DUBROVSKY ow often do new techREAL-TIME VS. nologies actually take us CONVENTIONAL PCR a step back? Usually, that is not the case, nor is it so with Real-time PCR, or quantitathe real-time PCR (also known tive PCR (qPCR), is a DNA-based as TaqMan), since its introducdetection assay where a PCR+ Real-time PCR is a demonstratively tion to the agricultural diagnostic amplified product is detected by more sensitive testing method than market in early 2000. Not only a fluorescing signal as it forms, in ELISA or conventional PCR. does the real-time PCR provide real-time, as opposed to conven+ Well-designed and optimized qPCR a more sensitive detection in tional PCR, where the product is assays can accurately detect the faster time, it also offers a less detected after completion of PCR presence of grapevine viruses. expensive testing alternative (for cycles by gel electrophoresis. + Because of its high specificity, qPCR example, assay costs in real-time Two main types of qPCR technology can be used for differenPCR usually cost less than ELISA chemistries exist: probe-based tiation of virus strain variants. or even traditional gel-based Ta q M a n a s s a y s a n d S Y B R PCR). Green assays, where fluoresc+ qPCR technology has the potential In response to the article ing, DNA-binding dye is used to be used in disease monitoring and ���Detecting Virus in Your Vines,��� instead of probe. In convenimplementation of pest management published in the Septembertional PCR, correctness of the strategies. October issue of this magaproduct is determined based on zine, we, as plant pathologists its size; in real-time PCR, it is at AL&L Crop Solutions Inc., tested more thoroughly: at the felt obligated to provide a more sequence level. in-depth view of this technology, its capabilities and In TaqMan assays, this is achieved by use of a applications, and to address some concerns and what probe, a short oligonucleotide, which binds on the I believe are misconceptions presented by the author. DNA strand between the primers. If the right target These include statements about the sensitivity of this sequence is found, a fluorescent signal is released technology and its ability to detect virus variants. from the probe during the product synthesis. If the AT A GLANCE 76 V I N E YARD & WINERY MANAGEMENT | Jan - Feb 2013 w w w. v w m m e d i a . c o m