Detection of toxic chemicals with high sensitivity by measuring the quantity of induced P450 mRNAs based on surface plasmon resonance.
In this study we describe a novel sensor system to detect toxic chemicals based on measurement of the quantity of Saccharomyces cerevisiae P450 mRNAs induced by them. Detection was conducted using a flow-injection-type sensor system based on surface plasmon resonance (SPR). The DNA and peptide nucleic acid (PNA) probes containing a complementary sequence to a part of P450 mRNA were immobilized on the sensor chip and the P450 mRNAs hybridized to the probes were quantified. We succeeded in detecting 10 ng/L (10 ppt) of atrazine using both DNA and PNA probes. Using this sensor system, we were able to detect bisphenol A in addition to atrazine. Furthermore, we achieved higher sensitivity by amplifying the target P450 mRNA based on nucleic acid sequence-based amplification (NASBA). This method allows for sensitive, rapid, and easy detection of some toxic chemicals.
Oyama
M
M
Research Center for Advanced Science and Technology, University of Tokyo, 4-6-1 Komaba, Meguro-ku, Tokyo 153-8904, Japan.
Ikeda
T
T
Lim
T
T
Ikebukuro
K
K
Masuda
Y
Y
Karube
I
I
D001280:Atrazine / Q000032:analysis*; D001483:Base Sequence; D002138:Calibration; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism*; D004247:DNA / Q000032:analysis; D017931:DNA Primers / Q000737:chemistry; D020135:Peptide Nucleic Acids / Q000737:chemistry; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000096:biosynthesis* / Q000737:chemistry; D012441:Saccharomyces cerevisiae / Q000737:chemistry*; D012965:Sodium Chloride / Q000737:chemistry; D020349:Surface Plasmon Resonance / Q000379:methods
D017931:DNA Primers; D020135:Peptide Nucleic Acids; D012333:RNA, Messenger; D012965:Sodium Chloride; D004247:DNA; D003577:Cytochrome P-450 Enzyme System; D001280:Atrazine