应用酶联免疫吸附试验检测马铃薯卷叶病毒

以辣根过氧化物酶标记马铃薯卷叶病毒抗体,采用双抗体夹心ELISA方法鉴定了马铃薯和洋酸浆的茎、叶、根及马铃薯块茎中的马铃薯卷叶病毒(Potato Leafroll Virus,PLRV),结果表明,对提纯的PLRV可测出的最低浓度为25ng/ml,当包被抗体浓度为40μg/ml、酶标记抗体稀释度为1/120时,可测出马铃薯茎、叶和根汁液中的PLRV,感染PLRV的洋酸浆茎、叶和根汁液的消光值,均比无病对照者高二倍以上,虽然感染PLRV的马铃薯休眠块茎维管束组织汁液的消光值高于无病毒对照,且脐部维管束组织消光值高于顶端,但测定打破休眠的感病块茎顶端维管束组织的阳性结果更为可靠和明显。

ENZYME-LINKED IMMUNOSORBENT ASSAY FOR THE DETECTION OF POTATO LEAFROLL VIRUS

The antibodies to PLRV were labelled with horse-radish peroxidase. The purified PLRV and PLRV in leaf, stem and root extracts of potato an Phy-salise floridana, as well as in vescular tissue of potato tubers were tested by direct double-sandwish assay of ELISA. The minimal amount of purified PLRV detected was 25ng/ml. The PLRV in leaf, stem and root extract were readily detected. Although the extinction values for PLRV in vascular tissue at heel end and rose end of dprmant tubers were higher than that of healthy tubers, it is more reliable to detect PLRV in rose end of tubers after breaking dormancy. The utilization of ELISA for certification of seed potatoes in China is discussed.