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Enterovirus_Genotyping
| Code No. | Description | Tests | Link |
| ENT-RR001 | Enterovirus RNA Real Time | 50 |  |
| ENT-RR002 | Enterovirus RNA Real Time | 100 | |
| ENT-RR003 | Enterovirus RNA Real Time | 200 |
ENTERO RNA CHECK consists of a primer/hybridization probe mix for the detection of enteroviral RNA with one step real-time RT-PCR*. In combination with commercially available kits (e.g. LC RNA Amplification Kit Hybridization Probes, Roche) enteroviral-RNA is detected with high specificity and sensitivity.
The kit is lyophilized and has to be reconstituted before use as described.
Principle
- The enteroviral specific amplicon is detected in real time by two distinctly labelled hybridization probes via fluorescence resonance energy transfer (FRET).
Specificity and sensitivity
- Poliovirus strains Sabin 1-3; Coxsackie A9, A7, B1; Echovirus 4, 5, 20 and 29. The sensitivity is determined with a pool of polio vaccines.
- In one RT-PCR* reaction 0,2 TCID50 are detected.
Content
- The kit is lyophilized and has to be reconstituted before use with 100µl H2OPCRgrade.
- The reconstituted primer/probe mix is stable for one week at 4°C or for six months at
-20°C. - After reconstitution it is recommended to store aliquots of the mix at -20°C. Multiple freeze-thaw cycles should be avoided.
Additional reagents not supplied
- RT-PCR*-Kit (e.g. LC RNA Amplification Kit Hybridization Probes, Roche).
Storage
- The kit has to be stored at 4°C in the dark.
- After reconstitution, the kit is stable up to one week at 4°C or for at least 6 months at -20°C.
- It is recommended to aliquote the vial to avoid frequent freezing and thawing.
- The kit has to be stored protected from light.
Assay procedure one step real-time RT-PCR*
- briefly spin the vial
- Reconstitution: add 100µl H2OPCRgrade, dissolve and mix (vortex); Attention:
please assure that the lyophilisate is completely dissolved. - Stability: The dissolved mix is stable at 4°C for one week or at –20°C for at least 6 months. Protect from light.
- Add 2µl of the Entero Check per 20µl RT-PCR reaction.
- ENTERO CHECK is applicable with commercially available Kits for RT.
- For the one-step real-time RT-PCR* in the LightCycler™ the "LC RNA Amplification
Kit Hybridization Probes" (Roche) is recommended. The kit has to be used as recommended by the supplier - H2OPCRgrade ads 20 µl ENTERO CHECK 2.0 µl 5x LC-RT-PCR* Mix 4.0 µl 25 mM MgCl2 0.8 µl RT-PCR* Enzyme-Mix 0.4 µl Template 1 - 10 µl
