Non-modified magnetic beads coupled with multiple real-time PCR for detection and quantification of mycotoxigenic fungi in paprika samples.
- Author:
Yan JIN
1
;
Wei-Wei ZHANG
;
Su-Yuan WANG
;
Zheng-Mao YE
;
Li-Shi ZHANG
;
Xiao-Fang PEI
Author Information
- Publication Type:Journal Article
- MeSH: Aspergillus; Capsicum; microbiology; DNA Primers; Food Contamination; analysis; Food Microbiology; Fungi; isolation & purification; Fusarium; Magnetic Phenomena; Penicillium; Real-Time Polymerase Chain Reaction; methods; Reproducibility of Results; Sensitivity and Specificity
- From: Journal of Southern Medical University 2015;35(1):23-28
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo 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).
METHODSThe 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.
RESULTSThe detection limit of this assay for spiked samples was 10(4) 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).
CONCLUSIONNMB-multiple qPCR assay we established allows rapid and sensitive detection of common mycotoxigenic fungi in paprika.
