Biotechnology and Bioengineering, Vol.71, No.3, 217-222, 2000
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.
Keywords:Saccharomyces cerevisiae;P450 14DM;atrazine;surface plasmon resonance (SPR);nucleic acid sequence-based amplification (NASBA)