Capillary Electrophoresis with Electrochemiluminescent Detection for Highly Sensitive Assay of Genetically Modified Organisms

Abstract

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.

Fluorescence aptasensor for Ochratoxin A in food samples based on hyperbranched rolling circle amplification

“…bind with the groove of dsDNA with high affinity and produce high fluorescence signal. 26 The fluorescence intensity of the system has some relationship with the OTA concentration, based on which, a sensitive fluorescence aptasensor can be developed. A simple assistant experiment has been performed to verify our presumption.…”

“…25 Since the products of HRCA consist of large amounts of double-stranded DNA (dsDNA) and singlestranded DNA (ssDNA) with various lengths. 26 SYBR Green I can bind with the groove of dsDNA with high affinity and 70 produce high fluorescence signal. High sensitivity fluorescence biosensors for protein based on HRCA had been reported 27,28 .…”

Characterization and Study of Transgenic Cultivars by Capillary and Microchip Electrophoresis

“…As can be seen in Table 1 , besides fluorescence detection, CGE has been employed in combination with other detection systems for the detection of PCR products [ 32 , 33 , 34 ]. Jiang et al reported a methodology for the determination of PCR amplified DNA fragments from transgenic soybeans using CE in its simplest mode (capillary zone electrophoresis, CZE) with UV detection [ 32 ].…”

Separation and determination of anesthetics by capillary electrophoresis with mixed micelles of sodium dodecyl sulfate and Tween 20 using electrochemiluminescence detection

“…Electrochemiluminescence (ECL) detection based on tris(2,2′–bipyridyl)ruthenium(II) chloride hexahydrate (Ru(bpy) 3 2+ ) has proved to be a powerful analytical technique due to its inherent features such as high sensitivity, low background, simple instrumentation and rapidity . Recently, CE combined with ECL (CE–ECL) has attracted much attention and has been extensively applied in the analysis of drugs , polymerase chain reaction (PCR) amplicon , amino acids and proteins .…”