( time PCR format which provides rapid a...)HSVision Molecular Beacon Detection Module Rapidly Detects Herpes Simplex,,,Virus DNA

Molecular Beacon Technology

Molecular beacons are single-stranded oligonucleotides that possess a stem-and-loop hairpin structure.^4-7 The loop is comprised of a DNA sequence complimentary to the target DNA, and the stem is formed by short complimentary sequences at opposite ends of the molecule. The ends of molecular beacons are labeled with a fluorophore and a quencher, respectively. When the molecular beacon is folded, the fluorophore and quencher are in close proximity, and fluorescence is quenched. However, when the molecular beacon is bound to a complimentary target, the fluorophore and quencher are separated, and the molecular beacon fluoresces (Figure 1).

Fig.1

Detecting Both HSV-1 and HSV-2 DNA

The HSVision module primers are derived from a 109 base-pair region of the gB gene that is highly conserved in HSV-1 and HSV-2 genomes.⁸ Because PCR reactions are often susceptible to a variety of inhibitors in biological specimens, an IAC is included in the HSV detection module to control for PCR failure. The IAC is a plasmid encoding a sequence identical in size and GC content to the HSV target DNA. The HSV-TC molecular beacon is labeled on the 5 end with the fluorophore tetrachloro 6-carboxyfluorescein (TET). The IAC-specific molecular beacon is labeled on the 5 end with 6-carboxyfluorescein (FAM). DABCYL is attached to the 3 end both molecular beacons and is used to quench fluorescence. The HSV-TC and IAC molecular beacons are multiplexed in a single-tube, real-time PCR reaction,
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