( allowing simultaneous detection of H...)HSVision Molecular Beacon Detection Module Rapidly Detects Herpes Simplex,,,Virus DNA

Amplification is performed with a PCR mixture that contains the HSV-TC and IAC molecular beacons, HSV PCR primers, dNTPS, Taq2000 DNA polymerase, an IAC DNA template, and an optimized PCR buffer. PCR is performed in a real-time spectrofluorometric thermal cycler. The presence of HSV-1 or HSV-2 DNA is determined by comparing the endpoint fluorescence value and/or the threshold cycle (Ct) values of the HSV DNA to the IAC control. As shown in Figure 2, 10⁵ to 10² copies of HSV-1 (Figure 2) and HSV-2 (Figure 2) are reliably detected with the HSV-TC molecular beacon.

Fig.2

Conclusions

Stratagenes single-tube HSVision molecular beacon detection module offers several advantages over existing HSV screening techniques: The test is performed in a closed tube so no post-PCR manipulation of samples is required. Because the entire reaction takes place in one tube, the test saves time and effort and significantly reduces the risk of PCR product carry-over contamination. Furthermore, this multiplexed assay includes both an HSV-specific molecular beacon as well as an internal control so assay performance and validation are realized. Hence, a true negative result can be discriminated from a false negative result due to PCR failure. In addition, the inclusion of both high- and low-copy number mixed HSV-1/HSV-2 DNA standards permits assay sensitivity to be validated. Using a 96-well format, samples can be completely screened in approximately 4 hours.

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2. Roizman, B. and Sears, A.E. (1996) In Fields Virology V.2, 3rd ed., (Eds. B.N., Knipe, D.M., and P.M Howley) Philadelphia Lippinco
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