MACRO: A Combined Microchip-PCR and Microarray System
for High-Throughput Monitoring of Genetically Modified Organisms
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Abstract
The monitoring of genetically modified
organisms (GMOs) is a primary
step of GMO regulation. However, there is presently a lack of effective
and high-throughput methodologies for specifically and sensitively
monitoring most of the commercialized GMOs. Herein, we developed a
multiplex amplification on a chip with readout on an oligo microarray
(MACRO) system specifically for convenient GMO monitoring. This system
is composed of a microchip for multiplex amplification and an oligo
microarray for the readout of multiple amplicons, containing a total
of 91 targets (18 universal elements, 20 exogenous genes, 45 events,
and 8 endogenous reference genes) that covers 97.1% of all GM events
that have been commercialized up to 2012. We demonstrate that the
specificity of MACRO is ∼100%, with a limit of detection (LOD)
that is suitable for real-world applications. Moreover, the results
obtained of simulated complex samples and blind samples with MACRO
were 100% consistent with expectations and the results of independently
performed real-time PCRs, respectively. Thus, we believe MACRO is
the first system that can be applied for effectively monitoring the
majority of the commercialized GMOs in a single test