Capillary electrophoresis with electrochemiluminescent detection for highly sensitive assay of genetically modified organisms.
A capillary electrophoresis coupled with electrochemiluminescent detection system (CE-ECL) was developed for the detection of polymerase chain reaction (PCR) amplicons. The ECL luminophore, tris(1,10-phenanthroline) ruthenium(II) (Ru(phen)(3)(2+)), was labeled to the PCR primers before amplification. Ru(phen)(3)(2+) was then introduced to PCR amplicons by PCR amplification. Eventually, the PCR amplicons were separated and detected by the homemade CE-ECL system. The detection of a typical genetically modified organism (GMO), Roundup Ready Soy (RRS), was shown as an example to demonstrate the reliability of the proposed approach. Four pairs of primers were amplified by multiple PCR (MPCR) simultaneously, three of which were targeted on the specific sequence of exogenous genes of RRS, and another was targeted on the endogenous reference gene of soybean. Both the conditions for PCR amplification and CE-ECL separation and detection were investigated in detail. Results showed that, under the optimal conditions, the proposed method can accurately identifying RRS. The corresponding limit of detection (LOD) was below 0.01% with 35 PCR cycles.
Guo
Longhua
L
Ministry of Education, Key Laboratory of Analysis and Detection for Food Safety, Fujian Provincial Key Laboratory of Analysis and Detection for Food Safety, Department of Chemistry, Fuzhou University, Fuzhou, 350002, China.
Yang
Huanghao
H
Qiu
Bin
B
Xiao
Xueyang
X
Xue
Linlin
L
Kim
Donghwan
D
Chen
Guonan
G
D017931:DNA Primers / Q000235:genetics / Q000378:metabolism; D004563:Electrochemistry; D019075:Electrophoresis, Capillary / Q000379:methods*; D005998:Glycine / Q000031:analogs & derivatives / Q000494:pharmacology; D008163:Luminescent Measurements; D030781:Organisms, Genetically Modified / Q000235:genetics; D009942:Organometallic Compounds / Q000378:metabolism; D010618:Phenanthrolines / Q000378:metabolism; D030821:Plants, Genetically Modified; D016133:Polymerase Chain Reaction; D013025:Glycine max / Q000187:drug effects / Q000235:genetics; D000097797:Glyphosate
D017931:DNA Primers; D005998:Glycine; D009942:Organometallic Compounds; D010618:Phenanthrolines; D000097797:Glyphosate; C034282:tris-(1,10-phenanthroline)ruthenium