Identification of PCR-amplified genetically modified organisms (GMOs) DNA by peptide nucleic acid (PNA) probes in anion-exchange chromatographic analysis

被引：9

作者：

Rossi, Stefano ^{[1
]}

Lesignoli, Francesca ^{[1
]}

Germini, Andrea ^{[1
]}

Faccini, Andrea ^{[1
]}

Sforza, Stefano ^{[1
]}

Corradini, Roberto ^{[1
]}

Marchelli, Rosangela ^{[1
]}

机构：

[1] Univ Parma, Dipartimento Chim Organ & Ind, I-43100 Parma, Italy

来源：

JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY | 2007年 / 55卷 / 07期

关键词：

PNA; PCR products; ion exchange; GMO; DNA; soy; maize; POLYMERASE-CHAIN-REACTION; SURFACE-PLASMON RESONANCE; REAL-TIME DETECTION; CYSTIC-FIBROSIS; POINT MUTATION; BIOSENSOR TECHNOLOGY; HYBRIDIZATION; RECOGNITION; MAIZE; CROPS;

D O I：

10.1021/jf062063s

中图分类号：

S [农业科学];

学科分类号：

09 ;

摘要：

PCR products obtained by selective amplification of transgenic DNA derived from food samples containing Roundup Ready soybean or Bt-176 maize have been analyzed by anion-exchange HPLC. Peptide nucleic acids (PNAs), oligonucleotide analogues known to bind to complementary single-stranded DNA with high affinity and specificity, have been used as specific probes in order to assess the identity of the peaks observed. Two different protocols were adopted in order to obtain single-stranded DNA: amplification with an excess of one primer or digestion of one DNA strand. The single-stranded DNA was mixed with the PNA probe, and the presence of a specific sequence was revealed through detection of the corresponding PNA:DNA peak with significantly different retention time. Advantages and limits of this approach are discussed. The method was tested with reference materials and subsequently applied to commercial samples.

引用

页码：2509 / 2516

页数：8

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