Chemiluminescent and colorimetric detection of a fluorescein-labelled probe and a digoxigenin-labelled probe after a single hybridization step.
The objective of the simple and fast method we describe is the simultaneous hybridization of two non-radioactive probes and their detection from the same blot, using two different systems. These two probes are synthesized by PCR: one is labelled with fluorescein and the other with digoxigenin. The former is detected by chemiluminescence and the latter by colorimetry. We applied this rapid and simple method to the specific detection of multiplex polymerase chain reaction products. We used the human herpes simplex virus HSV1 and HSV2 PCR models studied in our laboratory.
Chenal
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V
Laboratoire des Rickettsiales et Chlamydiales, Institut Pasteur, Paris, France.
Griffais
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R
D001483:Base Sequence; D003124:Colorimetry; D017931:DNA Primers / Q000737:chemistry; D015342:DNA Probes / Q000032:analysis* / Q000737:chemistry; D004076:Digoxigenin / Q000737:chemistry*; D005240:Feasibility Studies; D019793:Fluorescein; D005452:Fluoresceins / Q000737:chemistry*; D018259:Herpesvirus 1, Human / Q000235:genetics; D018258:Herpesvirus 2, Human / Q000235:genetics; D006801:Humans; D008163:Luminescent Measurements; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D016133:Polymerase Chain Reaction
D017931:DNA Primers; D015342:DNA Probes; D005452:Fluoresceins; D004076:Digoxigenin; D019793:Fluorescein