For Research Use Only. Not for use in diagnostic procedures. Others LightCycler HSV 1 2 Detection Kit RUO LightCycler® HSV 1/2 Detection Kit 3.5.5.3.6.10 Kit for the detection and differentiation of Herpes simplex virus type 1 and Herpes simplex virus type 2 DNA in research samples, using the LightCycler® Instrument. 03315177001 LC HSV 1/2 Detect. Kit (RUO) LightCycler HSV 1/2 Detection Kit 04038377019123 Reagents, kits 48 tests (4 x 12) Not Available false The LightCycler® HSV 1/2 Detection Kit is specifically adapted for real-time, on-line PCR in glass capillaries using the LightCycler® Instrument.Herpes simplex virus type 1 (HSV 1) and type 2 (HSV 2) are amplified with specific primers in a PCR reaction. The amplicon is detected by fluorescence using a specific pair of hybridization probes. The hybridization probes consist of two different short oligonucleotides that hybridize to an internal sequence of the amplified fragment during the annealing phase of the amplification cycle. One probe is labeled at the 5’end with LightCycler® Red 640-N-hydroxy-succinimide ester (Red 640-NHS ester), and to avoid extension, modified at the 3’-end by phosphorylation. The other probe is labeled at the 3’-end with fluorescein. After specific hybridization to the template DNA the two probes come in close proximity, resulting in fluorescence resonance energy transfer (FRET) between the two fluorophores. During FRET, fluorescein, the donor fluorophore, is excited by the light source of the LightCycler® Instrument, and part of the excitation energy is transferred to LightCycler® Red 640- NHS ester, the acceptor fluorophore. The emitted fluorescence of LightCycler ® Red 640-NHS ester is then measured by the LightCycler® Instrument.A Melting Curve analysis will be performed after the PCR run to differentiate positive samples in HSV 1 or HSV 2. The sequence specific probe labeled with LightCycler® Red 640-NHS ester will, due to a nucleotide polymorphism in the hybridization sequence between HSV 1 and HSV 2, show a type specific melting behaviour. Melting points for HSV 1 and HSV 2 are reproducible and significantly different, and allow clear determination of the HSV subtype.In enzyme-based amplification processes such as PCR, efficiency of the PCR process can be reduced by inhibitors that may be present in the unpurified sample material. In order to control for such substances that may interfere with PCR amplification, a kitspecific internal control (IC) has been added to the LightCycler® HSV 1/2 Detection Kit. The LightCycler® HSV 1/2 Internal Control is a synthetic double stranded DNA molecule with primer binding sites identical to the HSV target sequence, comprising a unique hybridization probe binding region that differentiates the IC from the target specific amplicon. The LightCycler® HSV 1/2 Internal Control is added to the lysed sample before the purification step and co-purified/amplified with the HSV DNA from the specimen in the same PCR reaction (Dual-color detection). en The LightCycler® HSV 1/2 Detection Kit provides primers and Hybridization Probes necessary for amplification and specific detection of HSV 1 and HSV 2, as well as, a control template DNA for reliable interpretation of results.The components of the kit are designed for detection and differentiation of HSV 1 and 2 with minimal handling steps. The enzyme components for amplification, buffers and control template are supplied as ready-to-use mixes.To ensure maximum reliability of the kit, the Internal Control (IC) will prevent misinterpretation of false negative results caused by inhibition of the amplification or caused by inefficient extraction. The IC is designed to be amplified with the same primers as the HSV DNA. The IC is detected in channel F3, whereas the HSV 1/2 DNA is detected in channel F2. The IC is spiked into the sample in a low concentration, in order not to decrease the dynamic range of the PCR reaction. In a HSV 1/2 negative sample, the IC must be positive to exclude inhibition or insufficient extraction. In HSV 1/2 DNA positive samples, the Internal Control may give a negative result.The kit allows the detection and differentiation of 48 samples in an average run size of 12 samples, including all the necessary reagents for running 2 positive control and 1 negative control reactions per run (64 reactions).Extraction reagents are not included in the kit.To ensure efficient and reliable detection/diffentiation of HSV 1/2 viral DNA, it is recommended to combine the LightCycler® HSV 1/2 Detection Kit with a standardized and automated extraction system, such as the MagNA Pure LC Instrument.Note: The LightCycler® HSV 1/2 Detection Kit was designed for use with the LightCycler® Instrument, and the performance can be guaranteed on the LightCycler® Instrument only. en The LightCycler® HSV 1/2 Detection Kit allows rapid detection and differentiation of HSV 1 and HSV 2 DNA and simultaneous detection of a kit specific internal control by dual-color detection. It is intended as a research tool for the qualitative detection and differentiation of HSV DNA in nucleic acid preparations from cell culture, swabs, plasma, serum and other biological samples.The LightCycler® HSV 1/2 Detection Kit was developed to be used in research applications such as studies on epidemiology, transmission or inactivation. It must not be used in diagnostic applications. en