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用于大规模诊断和分子鉴别从柑橘果园采集的温州蜜柑萎缩病毒的免疫层析检测试剂盒与双抗体夹心酶联免疫吸附测定法的比较

Comparison of an Immunochromatographic Assay Kit with DAS-ELISA for Large-Scale Diagnosis and Molecular Discrimination of Satsuma Dwarf Virus Collected from Citrus Orchards.

作者信息

Kato Mitsuhiro, Tomimura Kenta, Ishii Kanako

机构信息

Shizuoka Prefectural Research Institute of Agriculture and Forestry, Fruit Tree Research Center, Mobata, Shimizu, Shizuoka 424-0101, Japan.

Institute of Fruit Tree and Tea Science, National Agriculture and Food Research Organization (NARO), Okitsu, Shimizu, Shizuoka 424-0292, Japan.

出版信息

Plant Pathol J. 2020 Oct 1;36(5):509-514. doi: 10.5423/PPJ.NT.06.2020.0096.

Abstract

Satsuma dwarf virus (SDV) seriously damages citrus production by reducing the quality and yield of fruit. To avoid contamination with SDV, mother trees are checked to be SDV-free in advance of nursery tree distribution. In this study, we compared an immunochromatographic assay (ICA) kit with double-antibody sandwich enzyme-linked immunosorbent assay (DASELISA) for large-scale diagnosis of SDV in orchardgrown trees in Shizuoka Prefecture, Japan. The two methods gave conflicting results for 11 of 1,705 samples, all of which were negative by DAS-ELISA but positive by ICA. The samples scored as positive by either DASELISA or ICA were analyzed by reverse transcription polymerase chain reaction and all were confirmed to be positive. These results validate the use of ICA as a screening method for large-scale diagnosis. Strain discrimination revealed that 16 of 22 isolates belonged to SDV, while citrus mosaic virus (CiMV) infection only and co-infection (SDV and CiMV) were in a minority.

摘要

温州蜜柑萎缩病毒(SDV)通过降低果实品质和产量严重损害柑橘生产。为避免感染SDV,在分发苗木前会预先检查母树是否无SDV。在本研究中,我们比较了免疫层析检测(ICA)试剂盒和双抗体夹心酶联免疫吸附测定(DASELISA),用于对日本静冈县果园种植的树木进行大规模SDV诊断。在1705个样本中,有11个样本两种方法得出了相互矛盾的结果,所有这些样本通过DAS-ELISA检测为阴性,但通过ICA检测为阳性。通过DAS-ELISA或ICA检测为阳性的样本通过逆转录聚合酶链反应进行分析,所有样本均被确认为阳性。这些结果证实了ICA作为大规模诊断筛选方法的实用性。毒株鉴别显示,22个分离株中有16个属于SDV,而仅感染柑橘花叶病毒(CiMV)和同时感染(SDV和CiMV)的情况占少数。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e721/7542029/80e6d1c30607/PPJ-36-509-f1.jpg

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