Development of Rapid Detection Method for Cymbidium Mosaic Virus
Date Issued
2007
Date
2007
Author(s)
Chiou, Shih-Hao
DOI
zh-TW
Abstract
Phalaenopsis sp., an ornamental crop belonging to the Orchidacea, is an important commercial crop in the floral industry. Taiwan is one of the biggest exporters of Phalaenopsis orchid in the world market. Cymbidium Mosaic virus（CymMV）is one of the most economically important virus of Phalaenopsis orchid. The yield and quality of orchid production are influenced by this virus. To solve this problem, the virus-free orchids that used for breeding and the management of viral disease in orchid nurseries need to be developed. Therefore, a reliable and effective detection method of CymMV are needed. Enzyme-linked immunosorbent assay （ELISA）is a common method to detect CymMV in the orchid industry in Taiwan; however, high ratio of false positive detection are reported. Alternatively, reverse transcription polymerase chain reaction（RT-PCR）can be applied to the detection of CymMV, however, it is costy and highly trained technician are required for its accuracy. In this study, we found that low accumulation of some isolates of CymMV in orchids contribute partly to the high ratio of false positive detection of CymMV. Surprisingly, some isolates accumulate to high levels but still can’t be detected by ELISA. Sequence analysis revealed distinctive amino acids changed in the coat protein ORF of these isolates. To overcome the problems that applying RT-PCR in commercial CymMV detection, we alleviated the efforts of nucleic acids purification that commonly used in RT-PCR detection and develop a RT-PCR based CymMV rapid detection methods. The sensitivity of this method was similar to 3 commonly used RNA extraction methods in CymMV detection when same amounts of purified nucleic acid are used as template. Because the commonly used nucleic acid purification methods used for RT-PCR detection will cause nucleic acid loss (the yield loss range from 40.8% ~ 96%) during the purification, the rapid detection methods is the most sensitive methods in CymMV detection.
Subjects
蝴蝶蘭
東亞蘭嵌紋病毒
酵素連結免疫抗體法
多元抗體
反轉錄聚合酵素鍊鎖反應
Phalaenopsis sp.
Cymbidium mosaic virus
Enzyme-linked immunosorbent assay
Polyclonal antibody
Reverse transcription polymerase chain reaction
Type
other
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