Innovative Application of Fluorescent Microsphere Based Assay for Multiple GMO Detection

被引：21

作者：

Fantozzi, Anna ^{[1
,2
]}

Ermolli, Monica ^{[1
]}

Marini, Massimiliano ^{[1
,3
]}

Balla, Branko ^{[1
]}

Querci, Maddalena ^{[1
]}

Van den Eede, Guy ^{[1
]}

机构：

[1] Commiss European Communities, Biotechnol & GMOs Unit, Inst Hlth & Consumer Protect, DG Joint Res Ctr, I-2120 Ispra, Va, Italy

[2] Univ Basel, Dept Clin Biol Sci DKBW, Inst Biochem & Genet, Ctr Biomed, CH-4058 Basel, Switzerland

[3] Commiss European Communities, Reference Mat Unit, Inst Reference Mat & Measurements, B-2440 Geel, Belgium

来源：

FOOD ANALYTICAL METHODS | 2008年 / 1卷 / 01期

关键词：

GMO detection; epsps; p35S; Multiple assays; Fluorescent beads;

D O I：

10.1007/s12161-007-9005-0

中图分类号：

TS2 [食品工业];

学科分类号：

0832 ;

摘要：

An innovative multiple screening protocol allowing the detection of specific DNA sequences of p35S and epsps in GM soya flours simultaneously was developed. For the application of the Luminex xMAP technology, two different sets of fluorescent beads were cross-linked to the specific oligonucleotide probes previously amplified and labeled by polymerase chain reaction (PCR) in the presence of a biotinylated nucleotide. The detection of the amplification products bounded by the streptavidin-phycoeritrin conjugate was performed using the Luminex-100 instrument. The system allows identification of extremely low amounts of labeled targets as of 7.8 10(-4) nmol of p35S antiprobe with an associated value of repeatability relative standard deviation (RSDr) equal to 2.8. Parameters such as repeatability, reproducibility, limit of detection (LOD), and limit of quantification (LOQ) were considered to evaluate the performances of the assay. The potentiality of the system described in this paper enables us to look forward to a multiple target assay that is able to simultaneously detect and eventually quantify tens of target sequences occurring within the same sample preparation. The present study describes the first application of a multitarget fluorescent microsphere-based assay for DNA genetically modified organism (GMO) detection.

引用

页码：10 / 17

页数：8

共 41 条

[1]

*APPL BIOS, 2001, USERBULLETIN APPL BI, V2

[2] Oligogenic combinations associated with breast cancer risk in women under 53 years of age [J].