title: MACRO: a combined microchip-PCR and microarray system for high-throughput monitoring of genetically modified organisms.
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.
authors.affiliation: Shanghai Center for Systems Biomedicine, Key Laboratory of Systems Biomedicine (Ministry of Education), Shanghai Jiao Tong University , Shanghai 200240, China.
authors.forename: Ning
authors.initials: N
authors.lastname: Shao
authors.forename: Shi-Meng
authors.initials: SM
authors.lastname: Jiang
authors.forename: Miao
authors.initials: M
authors.lastname: Zhang
authors.forename: Jing
authors.initials: J
authors.lastname: Wang
authors.forename: Shu-Juan
authors.initials: SJ
authors.lastname: Guo
authors.forename: Yang
authors.initials: Y
authors.lastname: Li
authors.forename: He-Wei
authors.initials: HW
authors.lastname: Jiang
authors.forename: Cheng-Xi
authors.initials: CX
authors.lastname: Liu
authors.forename: Da-Bing
authors.initials: DB
authors.lastname: Zhang
authors.forename: Li-Tao
authors.initials: LT
authors.lastname: Yang
authors.forename: Sheng-Ce
authors.initials: SC
authors.lastname: Tao
mesh_terms: D017931:DNA Primers / Q000737:chemistry; D015342:DNA Probes / Q000737:chemistry; D018744:DNA, Plant / Q000235:genetics / Q000302:isolation & purification*; D057166:High-Throughput Screening Assays; D056656:Lab-On-A-Chip Devices; D057230:Limit of Detection; D018551:Solanum lycopersicum / Q000235:genetics; D046228:Microarray Analysis / Q000295:instrumentation / Q000379:methods*; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods*; D020411:Oligonucleotide Array Sequence Analysis / Q000295:instrumentation*; D012275:Oryza / Q000235:genetics; D030821:Plants, Genetically Modified; D013025:Glycine max / Q000235:genetics; D003313:Zea mays / Q000235:genetics
chemical_list: D017931:DNA Primers; D015342:DNA Probes; D018744:DNA, Plant