摘要
采用松材线虫的核糖体DNA的内转录区序列及cathepsin L-like cysteine proteinase为目的片段设计2对引物,建立了可特异性检测松材线虫的双基因PCR检测法。利用这2对引物,松材线虫可扩增出大小分别为490 bp及264 bp的产物,而其他对照组均无扩增。此检测方法经过实际应用检验,灵敏度与常规PCR一致,而特异性更高,可以应用于实验室的常规检测需求。
In this paper,a new two gene-jointed PCR detection assay to detect Bursaphelenchus xylophilus,was established.For developing the two gene-jointed PCR detection of B.xylophilus,two pairs of primers were designed according to sequence of B.xylophilus available in GenBank,targeting the cathepsin L-like cysteine proteinase gene and ribosomal DNA internal transcribed spacer region.The specificity,sensitivity assay and clinical samples were tested by using the optimized reaction system.Results showed that the specific fragments 490 bp and 264 bp were able to amplified,while there was no amplification product found in positive controls and other tested ones.The method was applied to detect clinical samples and the result showed 100 % consistence with PCR.These results could be served as a basis detection application in diagnosis of B.xylophilus.
作者
程维金
罗治建
陈亮
丁强
张叔勇
Chen Weijin;Luo Zhijian;Chen Liang;Ding Qiang;Zhang Shuyong(Wuhan Forestry Station,Wuhan 430023;Hubei Provincial General Station of Forest Pest Control and Quarantine,Wuhan 430079;Wuhan Baolvfeng Biotechnology Co.Ltd.,Wuhan 430048;Wuhan Institute of Virology,CAS,Wuhan 430071)
出处
《湖北林业科技》
2021年第2期60-62,65,共4页
Hubei Forestry Science and Technology
关键词
松材线虫
双基因PCR
检测方法
Bursaphelenchus xylophilus
two gene-jointed PCR
detection
