A rapid general method for the identification of PCR products using a fibre-optic biosensor and its application to the detection of Listeria.
A 200-mer fragment of the flaA gene from Listeria monocytogenes was amplified using the polymerase chain reaction (PCR) incorporating biotin- and fluorescein amadite (FAM-)labelled primers. Methods are described for isolating the single stranded FAM-labelled 200-mer. A central portion of this 200-mer was successfully hybridized onto a complementary sequence coated onto a fibre optic biosensor. Advantages in the specificity and speed of this approach compared to standard agarose gel electrophoresis and probing methods are discussed.
Strachan
N J
NJ
CSL Food Science Laboratory, Torry, MAFF, Aberdeen, UK.
Gray
D I
DI
D001426:Bacterial Proteins / Q000737:chemistry; D001483:Base Sequence; D015374:Biosensing Techniques; D001710:Biotin / Q000737:chemistry; D017931:DNA Primers / Q000235:genetics; D004269:DNA, Bacterial / Q000032:analysis*; D005336:Fiber Optic Technology; D005408:Flagellin / Q000235:genetics*; D019793:Fluorescein; D005452:Fluoresceins / Q000737:chemistry; D008089:Listeria monocytogenes / Q000302:isolation & purification*; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D016133:Polymerase Chain Reaction / Q000379:methods*; D019809:Streptavidin; D013997:Time Factors
D001426:Bacterial Proteins; D017931:DNA Primers; D004269:DNA, Bacterial; D005452:Fluoresceins; D005408:Flagellin; C067138:flaA protein, bacteria; D001710:Biotin; D019809:Streptavidin; D019793:Fluorescein