hygiena KIT230056 Norovirus GI GII plus Hepatitis A Virus Detection Kit User Guide
- June 2, 2024
- Hygiena
Table of Contents
hygiena KIT230056 Norovirus GI GII plus Hepatitis A Virus Detection Kit
Specifications
- Product: Norovirus (GI, GII) plus Hepatitis A Virus Detection Kit
- Model: KIT230056
- Revision : A, December 2023
- Application : PCR kit for the qualitative detection of hepatitis A virus and norovirus genogroup I and II
- Compatibility: Real-time PCR instruments
- Usage : For in vitro use only
- Kit Size: 64 reactions
Product Usage Instructions
Program Setup
Before setting up the PCR reactions, program your real-time PCR instrument as follows:
- Channels: u FAM (hepatitis A virus), HEX (norovirus GI), ROX (norovirus GII), and Cy5 (Process Control)
- Program Settings:
- 30 min, 1 cycle
- 5 min, 1 cycle
- 15 sec
- 60 sec*
- 10 sec, 50 cycles (*Fluorescence detection)
Data Interpretation
Verify the results of controls before interpreting sample results. Compare samples to positive and negative controls. Interpret results based on the table provided in the manual.
Preparation of the RT-PCR Mix
- PREPARE RT-PCR MIX: Mix carefully but thoroughly by pipetting up and down.
- ADD RT-PCR MIX.
- ADD SAMPLES AND CONTROLS.
- SEAL: Seal strips/plate accurately.
- CENTRIFUGE: Briefly spin strips/plate in a suitable centrifuge.
- START REAL-TIME PCR RUN: Cycle samples according to the programmed settings.
Note
This kit is intended for food testing purposes and is designed for the detection of hepatitis A virus and norovirus genogroup I and II.
FAQ
- Q : Can this kit be used for diagnostic purposes?
- A : This kit is intended for in vitro use only and is not suitable for diagnostic purposes. It is designed for qualitative detection in laboratory settings.
- Q : How should I store the kit when not in use?
- A : Store the kit according to the storage instructions provided in the product manual. Typically, it should be stored at the recommended temperature and protected from light.
PCR kit for the qualitative detection of hepatitis A virus and norovirus genogroup I and II using real-time PCR instruments. Before starting, it is strongly recommended to read the entire product manual available on our website.
PROGRAM SETUP
Program your real-time PCR instrument before setting up the PCR reactions. Select the following channels:
- FAM (hepatitis A virus), HEX (norovirus GI), ROX (norovirus GII) and Cy5 (Process Control).
- Reverse transcription: 1 cycle Step 1: 45 °C for 30 min
- Initial denaturation: 1 cycle Step 1: 95 °C for 5 min
- Amplifi cation 50 : cycles
- Step 1: 95 °C for 15 sec
- Step 2: 60 °C for 60 sec
- Step 3: 72 °C for 10 sec
- Fluorescence detection
For some real-time PCR instruments, the probe quencher as well as the usage of a passive reference dye has to be specified. This kit contains probes with a non-fluorescent “dark” quencher and no passive reference dye. For the LightCycler® 480 System, the generation of a colour compensation data file is necessary. Please use Color Compensation Set 3 (Product No. KIT230005).
DATA INTERPRETATION
Verify results of positive (Control Template) and negative controls (H2O), before interpreting sample results. Always compare samples to positive and negative control. Review data from each channel and interpret results as described in the table.
| FAM | HEX | ROX | Cy5 | Result Interpretation |
|---|---|---|---|---|
| + | + | + | + or – | Positive for hepatitis A virus, norovirus GI, norovirus GII |
| – | + | + | + or – | Positive for norovirus GI, norovirus GII |
| + | – | + | + or – | Positive for hepatitis A virus, norovirus GII |
| + | + | – | + or – | Positive for hepatitis A virus, norovirus GI |
| – | + | – | + or – | Positive for norovirus GI |
| + | – | – | + or – | Positive for hepatitis A virus |
| – | – | + | + or – | Positive for norovirus GII |
| – | – | – | + | Negative for hepatitis A virus, norovirus GI, norovirus GII |
| – | – | – | – | Invalid |
PREPARATION OF THE RT-PCR MIX
Take appropriate precautions to prevent contamination, e.g., by using filter tips and wearing gloves. Thaw reagents, mix (do not vortex!), and briefly spin vials before opening. Reverse transcriptase is a very temperature-sensitive enzyme. It is recommended to use a cooling block (at 4 °C) to avoid degradation of the Enzyme Solution (red cap). After usage, store the enzyme solution immediately at -15 to -25 °C.
- PREPARE RT-PCR MIX Add 14 μL of Master Mix (yellow cap) and 1 μL Enzyme Solution (red cap) for each reaction to a suitable tube (n samples + 2 controls + at least one additional reaction to cover pipetting loss). Mix carefully but thoroughly by pipetting up and down.
- ADD RT-PCR MIX Pipet 15 μL of prepared RT-PCR mix into each strip or plate well
- ADD SAMPLES AND CONTROL SPipet 10 μL of samples, Negative Control (orange cap) or Control Template (purple cap) into respective wells
- SEAL Seal strips/plate accurately.
- CENTRIFUGE Briefl y spin strips/plate in a suitable centrifuge.
- START REAL-TIME PCR RUN Cycle samples as described above
CONTACT
- foodproof® Norovirus (GI, GII) plus Hepatitis A Virus Detection Kit
- KIT230056 Kit for 64 reactions Store kit at -15 to -25 °C
- For food testing purposes FOR IN VITRO USE ONLY Made in Germany
- Hygiena®
- Camarillo, CA 93012 USA
- diagnostics.[email protected]
- www.hygiena.com