COMPARISON OF DOT-BLOT, ELISA, AND RT-PCR ASSAYS FOR DETECTION OF 2 CUCUMBER MOSAIC-VIRUS ISOLATES INFECTING BANANA IN HAWAII
COMPARISON OF DOT-BLOT, ELISA, AND RT-PCR ASSAYS FOR DETECTION OF 2 CUCUMBER MOSAIC-VIRUS ISOLATES INFECTING BANANA IN HAWAII
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DOI：
10.1094/pd-79-0902
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发表时间：
1995-09-01
期刊：
影响因子：
4.5
通讯作者：
JORDAN, R
中科院分区：
文献类型：
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作者：
HU, JS;LI, HP;JORDAN, R
对侵染夏威夷香蕉的两个黄瓜花叶病毒（CMV）分离物的外壳蛋白基因进行了克隆和序列测定。基于核苷酸和氨基酸序列的比较，这两个分离物属于CMV亚组I。一个分离物（CMV-夏威夷），其在夏威夷州的香蕉植物中是常见的并且诱导轻度花叶症状，与CMV-C株具有99%的序列同一性（核苷酸和氨基酸序列两者）。另一个分离物（CMV-Ostanus）仅在夏威夷州的两个香蕉农场发现，引起严重的花叶和叶片扭曲症状。CMV-Occurrence与CMV-C和CMV-Hawaii的核苷酸和氨基酸序列分别具有91%和93%的同一性。一种逆转录-聚合酶链反应（RT-PCR）检测香蕉样品中的两个CMV亚组。RT-PCR产物（约750 bp）也可作为探针用于斑点杂交检测CMV，PCR比斑点杂交和ELISA更敏感。斑点杂交法的灵敏度是ELISA法的100倍。CMV在香蕉植株内的分布不均匀。CMV浓度较高，在年轻的叶片比在老的，CMV-Occupy分离物没有检测到香蕉用抗体的CMV亚组I和II，但在斑点杂交和RT-PCR试验呈阳性。
The coat protein genes of two cucumber mosaic virus (CMV) isolates infecting banana plants in Hawaii were cloned and sequenced. Based on nucleotide and amino acid sequence comparisons, both isolates belong to CMV subgroup I. One isolate (CMV-Hawaii), which is common in banana plants in the state of Hawaii and induces mild mosaic symptoms, shares 99% sequence identity (both nucleotide and amino acid sequences) with CMV-C strain. Another isolate (CMV-Oahu), which was found only at two banana farms in the state of Hawaii, induces severe mosaic and leaf distortion symptoms. CMV-Oahu shares 91% and 93% nucleotide and amino acid sequence identity, respectively, with both CMV-C and CMV-Hawaii. A reverse transcription-polymerase chain reaction (RT-PCR) assay was developed for detection of both CMV subgroups from banana samples. The RT-PCR product (similar to 750 bp) was also labeled as a probe to detect CMV in dot blot hybridization tests, PCR is a more sensitive assay than either dot blot or ELISA. The dot blot assay was 100 times more sensitive than ELISA. The distribution of CMV within banana plants was uneven. CMV concentrations were higher in younger leaves than in older ones, The CMV-Oahu isolate was not detected in banana by ELISA using antibodies to both CMV subgroups I and II, but was positive in dot blot and RT-PCR tests.