Development of a real-time PCR-based method for detection of Xylophilus ampelinus

被引:14
|
作者
Dreo, T.
Gruden, K.
Manceau, C.
Janse, J. D.
Ravnikar, M.
机构
[1] Natl Inst Biol, Dept Plant Physiol & Biotechnol, Ljubljana 1000, Slovenia
[2] Jozef Stefan Inst, Ljubljana 1000, Slovenia
[3] Univ Angers, UMR Pathol Vegetale, INRA, INH,Ctr Angers, F-49071 Beaucouze, France
[4] Dept Bacteriol, Plant Protect Serv, NL-6706 EA Wageningen, Netherlands
关键词
grapevine bacterial blight; Vitis vinifera;
D O I
10.1111/j.1365-3059.2006.01487.x
中图分类号
S3 [农学(农艺学)];
学科分类号
0901 ;
摘要
A real-time PCR MGB-probe-based detection method specific to Xylophilus ampelinus, the cause of grapevine bacterial blight, was developed. Used in combination with the DNeasy plant mini kit, the sensitivity of X. ampelinus detection was approximately 100 cells from tissue extracts, surpassing the sensitivity of an existing nested PCR method at least tenfold. In field samples a high correlation was observed between real-time PCR cycle threshold (Ct) values obtained and X. ampelinus isolation on artificial media. Isolation was successful from samples with Ct values below 25. Lower concentrations of X. ampelinus, with Ct values up to 36, could also be reliably detected in real-time PCR. The newly developed method offers a reliable and sensitive test for X. ampelinus, suitable as a screening test, complementary to isolation on media or other methods, and could also be used for fast and specific identification of isolated colonies and for relative quantification of X. ampelinus bacteria.
引用
收藏
页码:9 / 16
页数:8
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