DEVELOPMENT OF A SIMPLE AND EFFECTIVE METHOD FOR SPECIFIC DETECTION OF PEPPER MILD MOTTLE VIRUS
文献类型： 外文期刊
作者： Wang, Y. 1 ; Li, X. 2 ; Liu, Y. 1 ; Wang, X. 1 ; Zhou, G. 1 ;
作者机构： 1.Chinese Acad Agr Sci, Inst Plant Protect, State Key Lab Biol Plant Dis & Insect Pests, Beijing 100193, Peoples R China
2.Beijing Acad Agr & Forestry Sci, Inst Plant & Environm Protect, Beijing, Peoples R China
关键词： Pepper mild mottle virus;digoxigenin-labeled cDNA probe;detection
期刊名称：ACTA VIROLOGICA （ 影响因子：1.162； 五年影响因子：1.255 ）
ISSN： 0001-723X
年卷期： 2009 年 53 卷 1 期
页码：
收录情况： SCI
摘要： A digoxigenin (DIG)-labeled cDNA probe complementary to the region from 5,256-6,300 nt of Pepper mild mottle virus (PMMoV) genome was synthesized. The specificity and sensitivity of the probe was tested by the (lot-blot hybridization. The detection limit of this method was equivalent to 0.8 mu g of fresh infected tissue in each spot. Double-antibody sandwich (DAS) ELISA and RT-PCR had the detection limit 39 mu g and 0.008 mu g of fresh infected tissue, respectively. We evaluated leaf, fruit pulp, and seed of pepper plant by dot-blot hybridization and found all the tested tissues Suitable for detection of PMMoV. Finally, 111 tissue samples including 93 samples collected from the pepper fields of Beijing and Baoding and 18 commercial seed samples Were evaluated by this method. The results showed that the incidence rate of the infected samples was 14% and 61% for the field samples and commercial seeds, respectively. The high sensitivity and reliability of the molecular hybridization assay provided an important alternative method for the detection of PMMoV in a large-scale.
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