ABSTRACT
A fully automated nucleic acid analysis system is described, which offers positive sample identification, improved sensitivity and reduced user interaction compared to conventional techniques. The system relies on the sequence-specific capture of DNA onto solid-phase particles, confirming product identity without the problems of interpretation and lack of sequence information inherent in gel-based analyses. The system can be used for sequence confirmation, mutation analysis and semiquantitative detection of PCR products.
Subject(s)
Nucleic Acids/analysis , Automation , Biotinylation , DNA Mutational Analysis/methods , DNA Primers , Microspheres , Oligonucleotides/analysis , Polymerase Chain Reaction , Sensitivity and Specificity , Sequence Analysis/methodsSubject(s)
Ethylene Oxide/administration & dosage , Proteins/drug effects , Carcinogens, Environmental/administration & dosage , Carcinogens, Environmental/metabolism , DNA/drug effects , DNA/metabolism , Dose-Response Relationship, Drug , Environmental Monitoring , Ethylene Oxide/metabolism , Gas Chromatography-Mass Spectrometry , Humans , Immunochemistry , Proteins/metabolismABSTRACT
A novel immunochemical approach has been developed to monitor human exposures to ethylene oxide (EO). The method exploits the interaction of EO with the amino function of the N-terminal valine residue of the alpha-chain of human haemoglobin (Hb). Antibodies were raised against the adducted valine in the form of the N-terminal tryptic heptapeptide and have been used to develop a sensitive radioimmunoassay (RIA) for the adducted heptapeptide. This method has been fully validated against a gas chromatography-mass spectrometry (GC-MS) method and has been applied to the monitoring of EO exposure in a group of sterilization workers.
