Harper Kathryn A, Smart Christine D, Davis R Michael
Department of Plant Pathology, University of California, Davis, CA 95616, USA.
J Forensic Sci. 2011 Jul;56(4):1003-9. doi: 10.1111/j.1556-4029.2011.01739.x. Epub 2011 Mar 10.
A DNA-based macroarray was designed to quickly and accurately identify certain Amanita mushroom specimens at the species level. The macroarray included probes for Amanita phalloides and Amanita ocreata, toxic species responsible for most mushroom poisonings, and Amanita lanei and Amanita velosa, edible species sometimes confused with toxic species, based on sequences of the highly variable internal transcribed spacer (ITS) region of rDNA. A cryptic species related to A. ocreata and one related to A. lanei, identifiable by ITS sequences, were also included. Specific multiple oligonucleotide probes were spotted onto nylon membranes and the optimal hybridization temperatures were determined. The Amanita DNA array was highly specific, sensitive (0.5 ng DNA/μL and higher were detected), and reproducible. In two case studies, the method proved useful when only small amounts of mushroom tissue remained after a suspected poisoning. An identification could be completed in 12 h.
设计了一种基于DNA的宏阵列，用于在物种水平上快速准确地鉴定某些鹅膏菌标本。该宏阵列基于核糖体DNA高度可变的内部转录间隔区（ITS）区域的序列，包含针对毒鹅膏和赭盖鹅膏（导致大多数蘑菇中毒的有毒物种）以及雷氏鹅膏和绒柄鹅膏（有时与有毒物种混淆的可食用物种）的探针。还包括了通过ITS序列可识别的与赭盖鹅膏相关的一个隐存种和与雷氏鹅膏相关的一个隐存种。将特定的多个寡核苷酸探针点样到尼龙膜上，并确定了最佳杂交温度。鹅膏菌DNA阵列具有高度特异性、灵敏性（可检测到0.5 ng DNA/μL及更高浓度）且可重复。在两个案例研究中，该方法在疑似中毒后仅残留少量蘑菇组织时被证明是有用的。鉴定可在12小时内完成。
