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本文报道了用点免疫印迹法(DIBA)检测单头桃蚜(Myzus persicae)和棉蚜(Aphis gossypii)所带黄 瓜花叶病毒(CMV)的试验结果,作者摸索出的DIBA法操作程序用于检测提纯的CMV最低下限浓度为0.16ng/ml,即每2μl样品含0.32pg抗原.这样的灵敏度是可以检测出单头蚜虫所带CMV的。这种操作程序是将蚜虫口针截取下来,浸入适当的缓冲液(2—5μl)内。这不仅保证了CMV病毒粒子充分地从蚜虫口针上释放出来.形成较高的浓度,而且极大限度地减少了非病毒物质对检测的干扰,因此就克服了从单头介体昆虫上检测所带植物病毒时常遇到的病毒浓度太低和非病毒抗源干扰两大困难。试验还表明。用这种DIBA法检出的蚜虫带毒率同生物测定试验检出的蚜虫传毒率有很好的一致性。显然,在蚜传非持久性病毒检测技术上的这一突破,对于植物病毒病流行学研究具有重要意义。
In this paper, we report the results of the DIBA method for the detection of cucumber mosaic virus (CMV) carried by Myzus persicae and Aphis gossypii by dot immunoblotting (DIBA) The lowest limit of detection of purified CMV was 0.16 ng / ml, which was 0.32 pg antigen per 2 μl of sample. This sensitivity allows the detection of single-aphid CMV. The procedure is to intercept the aphid mouth and immersed in the appropriate buffer (2-5 μl). This not only ensures that the CMV virions are adequately released from the aphids’ needles, but that higher concentrations are formed and the non-viral material is less susceptible to detection, thus overcoming detection from single-head insects The virus often comes with the plant virus concentration is too low and non-virus resistance to interference two major difficulties. Test also shows. The aphid infestation rate detected by this DIBA method is in good agreement with the aphid infestation rate detected in the bioassay. Obviously, this breakthrough in the detection of aphid-transmitted non-persistent virus is of great significance to the epidemiological study of plant virus disease.