摘要
以含马铃薯纺锤块茎类病毒(potato spindle tuber viroid,PSTV)RNA的总核酸为模板,加入人工合成的互补DNA引物,用反转录酶合成PSTV cDNA;在聚合酶链式反应系统中,用两个PSTV特异性引物进行cDNA扩增,用以制备光敏生物素标记的PSTV cDNA探针。用此探针进行斑点杂交检测含PSTV的马铃薯核酸提取液和汁液均出现阳性杂交信号,而健康马铃薯的核酸提取液和汁液的结果均为阴性。光敏生物素标记探针检测纯化PSTV的灵敏度可达5pg;检测感染PSTV的马铃薯块茎汁液的可测出最高稀释度为1:400。
Using potato spindle tuber viroid ( PSTV) -RNA-containinE total nucleic acid as template, PSTV-cDNA was synthesized by reverse transcription. The synthesized cDNA was then amplified by polymerase chain reaction ( PCR ) with two PSTV-specific primers. The amplified products were labelled with photobiotin to be used as probe for the detection of PSTV in infected potato tubers by dot-blot hybridization. The results showed that the PSTV either in partially purified RNA or in the extracted sap from diseased tubers was readily detected with strong positive signals,while the correspondent samples from heathy tuber gave negative reaction. The sensitivity of the probe for detecting the partially purified PSTV is 5 Pg.
出处
《病毒学报》
CAS
CSCD
北大核心
1992年第4期337-341,共5页
Chinese Journal of Virology
关键词
马铃薯类病毒
PSTV
聚合酶链反应
Potato spindle tuber viroid(PSTV) Polymerase chain reaction ( PCR ) Photobiotin-labelled cDNA probe Dot-blot hybridization
