Loncaric I, Donat C, Antlinger B, Oberlerchner J T, Heissenberger B, Moosbeckhofer R
Institute for Apiculture, Austrian Agency for Health and Food Safety (AGES), Vienna, Austria.
J Appl Microbiol. 2008 May;104(5):1433-41. doi: 10.1111/j.1365-2672.2007.03668.x. Epub 2007 Dec 7.
The aim of this study was to develop a specific and sensitive identification method for two Aureobasidium pullulans biocontrol strains, CF10 and CF40, based on a sequence-characterized amplified region (SCAR) derived from RAPD - and multiplex-RAPD PCR analysis.
The random amplified polymorphic DNA (RAPD) and multiplex RAPD-PCR techniques were used for a preliminary screening of A. pullulans genetic variability among 200 isolates. This approach allowed the selection of ten fragments present solely in strains CF10 and CF40. The RAPD fragments were cloned, sequenced and used to design two SCAR primers. Two primer pairs obtained from SCH3RAPD fragment of CF 40 and 6RAPD of CF10 were highly specific and sensitive.
In this study, we developed strain-specific multiplex-PCR based on sequence-characterized amplified region (SCAR) markers to simultaneously detect both strains in a single PCR.
This new multiplex-PCR provides a valuable tool for specific and sensitive identification of CF10 and CF40, and could be used in studies on the efficacy and persistence of introduced strains of A. pullulans for fire blight control.
本研究的目的是基于从随机扩增多态性DNA（RAPD）和多重RAPD-PCR分析中获得的序列特异性扩增区域（SCAR），开发一种针对两种出芽短梗霉生物防治菌株CF10和CF40的特异且灵敏的鉴定方法。
利用随机扩增多态性DNA（RAPD）和多重RAPD-PCR技术对200株出芽短梗霉的遗传变异性进行初步筛选。该方法筛选出了仅在菌株CF10和CF40中存在的10个片段。对RAPD片段进行克隆、测序，并用于设计两条SCAR引物。从CF40的SCH3RAPD片段和CF10的6RAPD获得的两对引物具有高度特异性和灵敏性。
在本研究中，我们基于序列特异性扩增区域（SCAR）标记开发了菌株特异性多重PCR，可在一次PCR中同时检测这两种菌株。
这种新的多重PCR为CF10和CF40的特异且灵敏的鉴定提供了一种有价值的工具，可用于研究引入的出芽短梗霉菌株对火疫病防治的效果和持久性。
