hygiena KIT230054 Hepatitis A Virus Detection Kit User Guide
- June 2, 2024
- Hygiena
Table of Contents
hygiena KIT230054 Hepatitis A Virus Detection Kit
Product Information
- Specifications
- Product Name: Hepatitis A Virus Detection Kit Ready Reference Guide
- Product No.: KIT230054
- Revision: A, December 2023
- Application: PCR kit for the qualitative detection of hepatitis A virus using real-time PCR instruments
- Usage: For food testing purposes
- Kit Size: 64 reactions
- For In Vitro Use Only
- Program Setup
- Before starting the PCR reactions, program your real-time PCR instrument with the following settings:
- Channels: FAM (hepatitis A virus) and ROX (Process Control)
- Program Steps:
- 30 min, 1 cycle
- 5 min, 1 cycle
- 15 sec
- 60 sec
- 10 sec, 50 cycles
- (Fluorescence detection)
- Data Interpretation
- Verify results of positive (Control Template) and negative controls (H2O) before interpreting sample results.
- Always compare samples to positive and negative controls.
- Interpret results based on the data from each channel as described in the table.
| FAM | ROX | Result Interpretation |
|---|---|---|
| + | – | Positive for hepatitis A virus |
| – | + or – | Negative for hepatitis A virus |
| + or – | – | Invalid |
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 as described in the program setup.
FAQs
- Q: Can this kit be used for diagnostic purposes?
- A: No, this kit is intended for food testing purposes only and is for in vitro use.
PROGRAM SETUP
Program your real-time PCR instrument before setting up the PCR reactions. Select the following channels: FAM (hepatitis A virus) and ROX (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.
DATA INTERPRETATION
Verify positive (Control Template) and negative control (H2 O) results 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 | ROX | Result Interpretation |
|---|---|---|
| + | + or – | Positive for hepatitis A virus |
| – | + | Negative for hepatitis A virus |
| – | – | 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. Using a cooling block (at 4 °C) is recommended 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 CONTROLS
- Pipet 10 µL of samples, Negative Control (orange cap) or Control Template (purple cap) into respective wells.
- SEAL
- Seal strips/plates accurately.
- CENTRIFUGE
- Briefly spin strips/plate in a suitable centrifuge.
- START REAL-TIME PCR RUN
- Cycle samples as described above.
- Product No. KIT230054
- Revision A, December 2023
- foodproof® is a registered trademark of Hygiena. The above-mentioned real-time PCR instruments are registered trademarks of their respective holders.
- foodproof®
- Hepatitis A Virus
- Detection Kit
- KIT230054
- 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.support@hygiena.com.
- www.hygiena.com.
References
Read User Manual Online (PDF format)
Read User Manual Online (PDF format) >>