Rezaian M A, Krake L R, Golino D A
CSIRO Division of Horticulture, Adelaide, Australia.
Intervirology. 1992;34(1):38-43. doi: 10.1159/000150261.
Pairs of viroid-specific oligonucleotide primers were selected and used in separate reverse transcription reactions coupled with the polymerase chain reaction to obtain DNA products of predetermined sizes characteristic of each viroid. The reaction conditions allowed efficient incorporation of small amounts of 32P-dATP which enabled rapid detection of the products in polyacrylamide gels. Using this method as well as probe hybridization, the presence of grapevine yellow speckle viroids 1 and 2 (previously known as GV1B) in grapevine samples from California was demonstrated, and it was established that the Australian grapevine viroid occurs in California. These comparisons provide the basis for uniform nomenclature of grapevine viroids found in different geographical regions.
选择了成对的类病毒特异性寡核苷酸引物，并将其用于与聚合酶链反应偶联的单独逆转录反应中，以获得每种类病毒特有的预定大小的DNA产物。反应条件允许有效掺入少量的32P-dATP，这使得能够在聚丙烯酰胺凝胶中快速检测产物。使用这种方法以及探针杂交，证明了加利福尼亚葡萄样品中存在葡萄黄斑类病毒1和2（以前称为GV1B），并且确定澳大利亚葡萄类病毒在加利福尼亚存在。这些比较为不同地理区域发现的葡萄类病毒的统一命名提供了基础。
