High sensitivity detection of 16s rRNA using peptide nucleic acid probes and a surface plasmon resonance biosensor.
A signal enhancing method allowing highly sensitive detection of E. coli 16s rRNA was developed using peptide nucleic acid (PNA) as a capture probe and a surface plasmon resonance (SPR) sensor as a detector. 16s rRNA has been used as a genetic marker for identification of organisms, and can be analyzed directly without PCR amplification due to the relatively high number of copies. PNA has a neutral backbone structure, therefore hybridization with 16s rRNA results in the ionic condition being changed from neutral to negative. A cationic Au nanoparticle was synthesized and used for signal amplification by ionic interaction with 16s rRNA hybridized on the PNA probe-immobilized SPR sensor chip. This method resulted in a detection limit of E. coli rRNA of 58.2+/-1.37 pg mL(-1). Using this analytical method, Staphylococcus aureus was detected without purification of rRNA.
Joung
Hyou-Arm
HA
BioNanotechnology Research Center, KRIBB, Daejeon, Republic of Korea.
Lee
Nae-Rym
NR
Lee
Seok Ki
SK
Ahn
Junhyoung
J
Shin
Yong Beom
YB
Choi
Ho-Suk
HS
Lee
Chang-Soo
CS
Kim
Sanghyo
S
Kim
Min-Gon
MG
D000818:Animals; D015374:Biosensing Techniques / Q000379:methods*; D004926:Escherichia coli / Q000737:chemistry; D053758:Nanoparticles / Q000737:chemistry; D009693:Nucleic Acid Hybridization / Q000379:methods; D015341:Nucleic Acid Probes / Q000378:metabolism; D020135:Peptide Nucleic Acids / Q000378:metabolism*; D012329:RNA, Bacterial / Q000032:analysis; D012336:RNA, Ribosomal, 16S / Q000032:analysis* / Q000378:metabolism; D012680:Sensitivity and Specificity; D013211:Staphylococcus aureus / Q000737:chemistry; D020349:Surface Plasmon Resonance
D015341:Nucleic Acid Probes; D020135:Peptide Nucleic Acids; D012329:RNA, Bacterial; D012336:RNA, Ribosomal, 16S