SIMULTANEOUS TD RT-PCR DETECTION OF CYMBIDIUM MOSAIC POTEXVIRUS AND ODONTOGLOSSUM RINGSPOT TOBAMOVIRUS WITH A SINGLE PAIR OF PRIMERS/

Cymbidium mosaic potexvirus (CymMV) and odontoglossum ringspot tobamov irus (ORSV) are the two most prevalent and economically important orch id viruses which have attained a world-wide distribution. We have desi gned one pair of common primers, which were able to detect, simultaneo usly, both viruses by a single polymerase chain reaction (PCR). Sequen ce alignment and primer analyses were employed in primer design. The e ffect of touch-down PCR, primer annealing temperature, number of therm al cycles, MgCl2 concentration and the degree of primer degeneracy, we re investigated and applied to the optimisation of PCR conditions in t he detection of both viruses in order to achieve equal amplification e fficiencies. This is the first successful attempt at using a single pa ir of PCR primers to detect two viruses belonging to different taxonom ic groups simultaneously. (C) 1998 Elsevier Science B.V. All rights re served.