MACRO: a combined microchip-PCR and microarray system for high-throughput monitoring of genetically modified organisms

Anal Chem. 2014 Jan 21;86(2):1269-76. doi: 10.1021/ac403630a. Epub 2014 Jan 9.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • DNA Primers / chemistry
  • DNA Probes / chemistry
  • DNA, Plant / genetics
  • DNA, Plant / isolation & purification*
  • Glycine max / genetics
  • High-Throughput Screening Assays
  • Lab-On-A-Chip Devices*
  • Limit of Detection
  • Microarray Analysis / instrumentation
  • Microarray Analysis / methods*
  • Multiplex Polymerase Chain Reaction / methods*
  • Oligonucleotide Array Sequence Analysis / instrumentation*
  • Oryza / genetics
  • Plants, Genetically Modified
  • Solanum lycopersicum / genetics
  • Zea mays / genetics

Substances

  • DNA Primers
  • DNA Probes
  • DNA, Plant