南方医科大学学报Issue(1)：23-28,6.DOI:10.3969/j.issn.1673-4254.2015.01.05
辣椒中常见产毒性真菌的裸磁珠-多重实时荧光定量PCR检测方法
Non-modified magnetic beads coupled with multiple real-time PCR for detection and quan-tification of mycotoxigenic fungi in paprika samples
摘要
Abstract
Objective To establish a method for detecting 3 common toxigenic molds (Aspergillus, Penicillium, and Fusarium) based on non-modified magnetic beads coupled with multiple real-time PCR (NMB-multiple qPCR). Methods The primers and genus-specific probes were designed based on the rDNA sequences to develop a multiple real-time PCR using non-modified magnetic bead to enrichment of fungal spores. The sensitivity, specificity and repeatability of this assay were evaluated. Results The detection limit of this assay for spiked samples was 104 CFU/g, demonstrating a 10-fold greater detection sensitivity of this assay than that of real-time PCR. The NMB-multiple qPCR assay also showed good specificity and reproducibility and yielded comparable results with those by traditional colony counting method for spiked samples (P>0.05). Conclusion NMB-multiple qPCR assay we established allows rapid and sensitive detection of common mycotoxigenic fungi in paprika.关键词
产毒性真菌/裸磁珠富集/多重实时荧光定量PCR/辣椒模型Key words
mycotoxigenic fungi/enrichment of non-modified magnetic beads/multiple real-time PCR/paprika model引用本文复制引用
金燕,张薇薇,王溯源,叶正茂,张立实,裴晓方..辣椒中常见产毒性真菌的裸磁珠-多重实时荧光定量PCR检测方法[J].南方医科大学学报,2015,(1):23-28,6.基金项目
国家自然科学基金（81030053）Supported by National Natural Science Foundation of China (81030053) （81030053）
