Non-modified magnetic beads coupled with multiple real-time PCR for detection and quan-tification of mycotoxigenic fungi in paprika samples

VernacularTitle: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
Author: Yan JIN ¹ ; Weiwei ZHANG ; Suyuan WANG ; Zhengmao YE ; Lishi ZHANG ; Xiaofang PEI
Author Information

1. 四川大学华西公共卫生学院卫生检验与检疫学系
Keywords: mycotoxigenic fungi; enrichment of non-modified magnetic beads; multiple real-time PCR; paprika model
From: Journal of Southern Medical University 2015;(1):23-28
CountryChina
Language:Chinese
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.