Purification and Antiserum Preparation of Maize Dwarf Mosaic Virus

玉米矮化嵌紋病毒之純化與抗血清製作

鄧汀欽(T. C. Deng) ； 黃秋雄(C. H. Huang)

《中華農業研究》 35卷3期 (1986/09) Pp. 350-359

https://doi.org/10.29951/JARC.198609.0010

摘要

利用等密度平衡離心法純化玉米矮化嵌紋病毒B型系統，以提高病毒純度及產量。純化所得之MDMV-B仍具相當之感染力，且純化過程並無病毒聚集現象。以紫外光吸收光譜測定病毒純度，可得一標準的核蛋白吸收曲線，其最高值在259nm最低值在249nm。純化的病毒產量爲每克葉片有70-80微克。利用純化的MDMV-B對白兔行肌肉注射以製作抗血清，其力價以瓊脂擴散反應測定最高爲1/8，但以沈澱環反應測定則爲1/2048。此等抗血清完全不與健康的玉米株汁液反應，可見利用本方法已獲得良好純化效果。從甘蔗分離出來而在玉米上繁殖的一個甘蔗嵌紋病毒系統與MDMV-B抗血清有反應，但是在瓊脂擴散反應試驗時形成刺狀沈澱帶，由此可以區分MDMV-B和本省之SCMV系統。另由酵素結合抗體免疫分析的結果，MDMV-B、SCMV-B及SCMV-D對MDMV-B抗體反應的讀值，同樣可以確認其間的異同。

並列摘要

A high purity of maize dwarf mosaic virus strain B (MDMV-B) with sufficient yield was obtained by isopycnic centrifugation in cesium sulphate. The virus was infectious and nonaggregated after the purification process. Ultraviolet absorption spectrum of the purified virus showed a typical curve of nucleroprotein with a maximum at 259 and a minimum at 249 nm. The yield of purified virus was 70-80μg per g of tissue. Antiserum to MDMV-B was produced by intramuscular injection of the purified virus to a rabbit. The highest titer of the antiserum obtained 7 weeks after the first injection was 1/8 in SDS-immunodiffusion test, and 1/2048 in precipitin ring test. The completely negative reactions of MDMV-B antiserum to healthy plant extract indicated the efficiency of this method for the purification of MDMV-B. A spur was produced between antigens of an isolate of sugarcane mosaic virus (SCMV) and MDMV-B against antiserum of MDMV-B in SDS-immunodif fusion test. Using the antiserum prepared in this study, we could easily distinguish MDMV-B from strains of SCMV by either SDS-immunodiffusion test or ELISA.