Wetzel T, Tavert G, Teycheney P Y, Ravelonandro M, Candresse T, Dunez J
Station de Pathologie Végétale, Institut National de la Recherche Agronomique, Centre de Bordeaux, Villenave d'Ornon, France.
J Virol Methods. 1990 Nov;30(2):161-71. doi: 10.1016/0166-0934(90)90017-a.
A cDNA library covering the complete genome of plum pox virus strain D (PPV D) has been obtained, and an endonuclease restriction map derived from it. This map was superposed on the PPV genomic organisation map, established for a nonaphid transmissible strain of PPV (Maiss et al., 1989). This allowed us to select seven probes, corresponding to different regions on the PPV genome. These probes were tested in a dot-blot hybridization assay for the detection of PPV. Probes of various lengths (0.25 to 1.5 kb) were tested and those measuring at least 0.8 kb (4 of the 7 probes selected) proved to be the most sensitive. The detection limit was of about 5 pg of purified virus per assay. Probes representing non-structural viral protein genes were equally sensitive in detecting both serotypes D and M of PPV. The previously described probe pBPPV1 (Varveri et al., 1988), covering the coat protein gene of strain D, was less sensitive, when compared to the above probes, in detecting heterologous strains of PPV. The polyvalence of probes transcribed from non-structural viral protein genes was confirmed by screening isolates of PPV, collected in infected orchards in several Mediterranean countries.
已获得一个覆盖李痘病毒D株（PPV D）完整基因组的cDNA文库，并由此得到了一个核酸内切酶限制图谱。该图谱与为PPV的一个非蚜虫传播株构建的PPV基因组组织图谱（Maiss等人，1989年）重叠。这使我们能够选择七个对应于PPV基因组不同区域的探针。这些探针在斑点杂交试验中用于检测PPV。测试了各种长度（0.25至1.5 kb）的探针，结果表明至少0.8 kb长的探针（所选的7个探针中的4个）最为灵敏。每次试验的检测限约为5 pg纯化病毒。代表非结构病毒蛋白基因的探针在检测PPV的D和M血清型时同样灵敏。与上述探针相比，先前描述的覆盖D株外壳蛋白基因的探针pBPPV1（Varveri等人，1988年）在检测PPV的异源株时灵敏度较低。通过对从几个地中海国家受感染果园收集的PPV分离株进行筛选，证实了从非结构病毒蛋白基因转录的探针具有多价性。
