摘要
松茸(Tricholoma matsutake)是一种稀有、珍贵的野生食用菌,至今还无法人工栽培,目前菌菇市场中存在大量假冒松茸商品。文章建立了快速鉴定松茸及其制品的可视化核酸试纸条法。该方法设计特异性引物,通过聚合酶链式反应(Polymerase Chain Reaction,PCR)扩增特异性鉴定松茸特异性基因标志物,优化扩增条件为退火温度63 ℃、15个循环。扩增产物使用胶体金核酸试纸条检测,松茸样本均出现2条红色条带,同时凝胶电泳验证可扩增出352 bp特异性DNA片段。其他菌类及空白对照均在质控线出现1条红色条带,检测线无显色条带,且凝胶电泳无扩增条带,结果特异良好。该方法灵敏度高、稳定性强,模板DNA的最低检出量为0.1 ng/μL,检测试纸条可在常温保存至少12个月。所建立的松茸特异性基因标志物成分检测的可视化核酸试纸条方法检测性能优异,结果稳定可视、操作简便,适用于松茸原料和加工制品的现场检测。
Tricholoma matsutake is a rare and precious wild edible mushroom,which cannot be cultivated artificially so far.At present,there is a large number of counterfeit Tricholoma matsutake products in the mushroom market.A visual nucleic acid test strip method for rapid identification of Tricholoma matsutake and its products was established.Specific primers were designed to amplify Tricholoma matsutake-specific genetic markers via polymerase chain reaction (PCR).The optimized amplification conditions were an annealing temperature of 63 ℃ for 15 cycles.The amplified products were detected using colloidal gold nucleic acid test strips.Tricholoma matsutake samples showed two red bands,and gel electrophoresis confirmed the amplification of a 352 bp specific DNA fragment.Other fungi and blank controls showed one red band on the control line without any color band on the test line,and no amplification bands in gel electrophoresis,demonstrating good specificity.This method exhibits high sensitivity and stability,with the lowest detectable amount of template DNA being 0.1 ng/μL.The test strips can be stored at room temperature for at least 12 months.The visual nucleic acid test strip method developed in study for detecting Tricholoma matsutake-specific genetic markers has excellent performance,providing stable,visible results,and is easy to operate,making it suitable for on-site detection of Tricholoma matsutake raw materials and processed products.
作者
朱建宇
尹荣
王艳双
常明珠
周亭亭
ZHU Jianyu;YIN Rong;WANG Yanshuang;CHANG Mingzhu;ZHOU Tingting(Basic Medicine College of Beihua University,Jilin 132013,China;Science College of Beihua University,Jilin 132013,China;Medical Technology College of Beihua University,Jilin 132013,China)
出处
《食品科技》
CAS
北大核心
2024年第5期342-348,共7页
Food Science and Technology
基金
2021年吉林省科技厅项目(20210203054SF)。
关键词
松茸
快速鉴定
聚合酶链式反应
核酸试纸条
可视化检测
Tricholoma matsutake
rapid identification
polymerase chain reaction
nucleic acid test strip
visual detection
