hygiena KIT230110 (LP) foodproof Clostridium botulinum Detection LyoKit User Guide
- May 15, 2024
- Hygiena
Table of Contents
hygiena KIT230110 (LP) foodproof Clostridium botulinum Detection LyoKit
Specifications
- Product Name: Clostridium botulinum Detection LyoKit
- Revision: A, November 2023
- Product Numbers: KIT230110 (LP), KIT230111 (RP)
- Functionality: PCR kit for the qualitative detection of botulinum type A, B, E, and F neurotoxin/producing Clostridium species
- Compatibility: Real-time PCR instruments
- Manufactured: Germany
Product Usage Instructions
Program Setup
- Program your real-time PCR instrument before setting up the PCR reactions. Select the following channels:
- FAM (botulinum neurotoxin types A and E)
- HEX (botulinum neurotoxin types B and F)
- ROX (Internal Control)
2. Follow the specified cycle settings provided in the manual.
Data Interpretation
- Verify results of positive (Control Template) and negative (H2O) controls before interpreting sample results.
- Compare sample results to positive and negative controls.
- Review data from each channel and interpret results based on the provided table.
Preparation of the PCR Mix
- Take appropriate precautions to prevent contamination by using filter tips and wearing gloves.
- Place strips in a rack and ensure lyophilized pellets are at the bottom of all tubes.
- Add samples and controls, then carefully seal the tubes with the provided caps.
- Mix thoroughly by resuspending the pellet either by gentle tapping or pipetting.
- Briefly centrifuge the strips to ensure proper mixing.
- Start the real-time PCR run as described in the manual.
For Food Testing Purposes:
This product is for in vitro use only. Follow all instructions carefully for accurate results.
Made in Germany
Frequently Asked Questions (FAQ)
Q: Can this kit differentiate between botulinum neurotoxin types A and E, and B and F?
- A: Without a melting curve analysis, the botulinum neurotoxin types can still be detected, but differentiation between A and E or B and F is not possible.
Q: What precautions should I take during the preparation of the PCR mix?
- A: Take appropriate precautions to prevent contamination by using filter tips, wearing gloves, and ensuring proper sealing of tubes.
- Product No. KIT230110 (LP), KIT230111 (RP)
- PCR kit for the qualitative detection of botulinum type A, B, E, and F neurotoxin/producing Clostridium species (C. botulinum, C. baratii, and C. butyricum) using real-time PCR instruments.
- Without a melting curve analysis, the botulinum neurotoxin types (BoNT) can still be detected but not differentiate between A and E or B and F.
- 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 (botulinum neurotoxin types A and E), HEX (botulinum neurotoxin types B and F), and ROX (Internal Control).
- Pre-incubation : 1 cycle
- Step 1: 37 °C for 4 min
- Step 2: 95 °C for 5 min
- Amplification: 50 cycles
- Step 1: 95 °C for 5 sec
- Step 2: 60 °C for 60 sec
- Melting Curve: 1 cycle
- Step 1: 95 °C for 50 sec
- Step 2: 37 °C for 50 sec
- For some real-time PCR instruments, the probe quencher as well as the use of a passive reference dye must be specified.
- This kit contains probes with a non-fluorescent “dark” quencher and no passive reference dye.
- A Color Compensation is necessary for users of the LightCycler® 480 System: Color Compensation Set 3 (Product No. KIT230005).
DATA INTERPRETATION
- Verify the results of positive (Control Template) and negative (H2O) controls, before interpreting the sample results.
- Always compare samples to positive and negative controls.
- Review data from each channel and interpret results as described in the table.
Amplification Curve| FAM| HEX| ROX| Result
Interpretation
---|---|---|---|---
+| –| + or –| Positive for BoNT A and/or E
–| +| + or –| Positive for BoNT B and/or F
+| +| + or –| Positive for BoNT A and/or E, Positive for BoNT B and/or F
–| –| + or –| Negative for BoNT A, B, E, F
–| –| –| Invalid
Melting Curve| FAM| HEX| Botulinum Neurotoxin Type (BoNT)
none| none| A
none| none| B
68 ± 2 °C| none| E
none| 55 ± 2 °C and 64.5 ± 2 °C| F
(64 ± 2 °C)| 55 ± 2 °C and 62.5 ± 2 °C| Control Template
PREPARATION OF THE PCR MIX
- Take appropriate precautions to prevent contamination, e.g., by using filter tips and wearing gloves.
- PLACE STRIPS IN THE RACK
- Take the needed number of PCR tube strips out of an aluminum bag.
- Important: close the bag tightly afterward. Place strips in a suitable PCR tube rack.
- If needed, gently tap the tubes to move the lyophilized pellets to the bottom of all tubes.
- DECAP
- Immediately before filling, carefully open strips and discard caps.
- Do not leave it open longer than necessary.
- ADD SAMPLES AND CONTROLS
- Pipette 25 µL of samples, Negative Control (colorless cap), or Control Template (purple cap) into respective wells.
- If using less volume, add PCR-grade H2O to reach 25 µL.
- SEAL
- Carefully seal the tubes with the provided 8-cap strips.
- MIX
- Resuspend the pellet after sealing by mixing thoroughly.
- Alternatively, resuspend the pellet by pipetting up and down multiple times in Step 3.
- CENTRIFUGE
- Briefly spin strips, e.g., 5 seconds at 500 – 1,000 x g, in a suitable centrifuge.
- START REAL-TIME PCR RUN
- Cycle samples as described above.
- Place tubes in a vertical, balanced order into the cycler, e.g., two strips can be placed in the first and last column.
- foodproof® Clostridium botulinum Detection LyoKitKIT230110 /11
- Kit for 96 reactions Store kit at 2 to 8 °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
Documents / Resources
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hygiena KIT230110 (LP) foodproof Clostridium botulinum Detection
LyoKit
[pdf] User Guide
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foodproof Clostridium botulinum Detection LyoKit, Clostridium botulinum
Detection LyoKit, botulinum Detection LyoKit, Detection LyoKit, LyoKit
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