高效价甘薯羽状斑驳病毒抗血清的制备

Preparation of High Titer Antiserum to Sweet Potato Feathery Mottle Virus(SPFMV)

用嫁接方法将甘薯羽状斑驳病毒（ＳＰＦＭＶ）接种到Ｉ．ｓｅｔｏｓａ上扩繁，以０．２ｍｏｌ／ＬｐＨ７．２ＰＢＫ缓冲液、垫层差速离心、蔗糖密度梯度离心提取纯化ＳＰＦＭＶ。纯化的ＳＰＦＭＶＯＤ２６０／２８０的比值为１．２５。将纯化的ＳＰＦＭＶ免疫家兔制备抗血清，在环状沉淀和微量沉淀试验中，用提纯病毒测定抗血清的效价均为１：４０９６；以ＳＰＦＭＶ－ＩｇＧ为第一抗体，应用Ｄｏｔ－ＥＬＩＳＡ对甘薯和Ｉ．ｓｅｌｏｓａ叶片中的ＳＰＦＭＶ分别作了测定。

weet potato feathery mottle virus(SPFMV)was inoculated to I.selosa to propagare by graftingand was purified wirh 0.2mol/L pH7.2 PBK.Further purification of SPFMV was conducted by sucrose cushion and sucrose density gradient centrifugation.Purified SPFMV had a OD260/280 ratio of1.25.Rilbbit was immuned with purifled viruses to prepare antiSerum.Antisera produced to purifiedvirus had a titer of 1:1096 in microprecipitin and ring precipitin tests.SPFMV in leaves of sweet potato and I.selosa were detected respectively by indirect enzyme-linked immunosorbent assay(ELISA)onnitreeellulose membranes.