Application of peptide nucleic acid to the direct detection of deoxyribonucleic acid amplified by polymerase chain reaction.
Double-stranded DNA amplified by polymerase chain reaction (PCR) was detected by peptide nucleic acid (PNA) using a BIAcore 2000 biosensor based on surface plasmon resonance (SPR). PNA is an artificial oligo amide that is capable of forming highly stable complexes with complementary oligonucleotides. We succeeded in the direct detection of double-stranded DNA, amplified by PCR with high-sequence specificity. It was shown that the target DNA was available for detection over the range of 40-160 nM. Therefore, the detection limit was 7.5 pmol of the target DNA (143 bases, applied volume 30 microliters). Our DNA detection system, the combination of BIAcore and the probe PNA, could detect the target DNA with good reproducibility. In this report, we show that our system is a powerful tool for the diagnosis of pathologically significant DNA.
Sawata
S
S
Research Center for Advanced Science and Technology, University of Tokyo, Japan.
Kai
E
E
Ikebukuro
K
K
Iida
T
T
Honda
T
T
Karube
I
I
D001427:Bacterial Toxins / Q000235:genetics*; D001483:Base Sequence; D015374:Biosensing Techniques; D004247:DNA / Q000032:analysis*; D004926:Escherichia coli; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D020135:Peptide Nucleic Acids; D016133:Polymerase Chain Reaction; D022641:Shiga Toxin 2; D020349:Surface Plasmon Resonance
D001427:Bacterial Toxins; D020135:Peptide Nucleic Acids; D022641:Shiga Toxin 2; D004247:DNA