hygiena KIT230088 (LP) Listeria Genus Detection LyoKit User Guide

hygiena KIT230088 (LP) Listeria Genus Detection LyoKit

Product Information

Specifications:

• Product Name: Listeria Genus Detection LyoKit
• Revision: A, November 2023
• Product Numbers: KIT230088 (LP), KIT230089 (RP), KIT230330 (DP)
• Application: PCR Kit for the qualitative detection of Listeria sensu stricto species
• Species Detected: L. monocytogenes, L. seeligeri, L. ivanovii, L. welshimeri, L. innocua, and L. marthii

Product Usage Instructions

Program Setup:

1. Program your real-time PCR instrument before setting up the PCR reactions.
2. Select the following channels: FAM (Listeria sensu stricto) and VIC (Internal Control).
3. Set program cycle as follows:
   • 4 min at 5 min for 1 cycle
   • 5 sec at 60 sec for 50 cycles
   • *Fluorescence detection

Data Interpretation:

1. Verify results of positive (Control Template) and negative controls (H2O) before interpreting sample results.
2. Always compare samples to positive and negative controls.
3. Review data from each channel and interpret results based on the provided table.

Preparation of the PCR Mix:

1. Take precautions to prevent contamination by using filter tips and wearing gloves.
2. Place strips in a rack, ensuring lyophilized pellets are at the bottom of all tubes.
3. Open strips just before filling, add samples and controls, then seal the tubes with provided caps.
4. Mix thoroughly to resuspend pellet and centrifuge briefly.
5. Start the real-time PCR run according to the programmed cycle.

FAQ:

• Question: What should I do if my results show an invalid interpretation?
  Answer: In case of an invalid interpretation, ensure proper handling of samples, controls, and following the PCR setup instructions accurately. If the issue persists, contact customer support for further assistance.

Product No. KIT230088 (LP), KIT230089 (RP), KIT230330 (DP)

PCR Kit for the qualitative detection of the Listeria sensu stricto species (L. monocytogenes, L. seeligeri, L. ivanovii, L. welshimeri, L. innocua and L. marthii).
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 (Listeria sensu stricto) and VIC (Internal Control).

As an alternative to VIC, HEX can be used. For the PikoReal® 24, Yakima Yellow has to be selected.

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 Dualo 32 R2 real-time PCR instrument, please open the software, click on ‘New’, and select the respective template file. Template files can be added by clicking on ‘Add’ in the ‘Select template file’ window.

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.

PREPARATION OF THE PCR MIX

Take appropriate precautions to prevent contamination, e.g., by using filter tips and wearing gloves.

1. PLACE STRIPS IN RACK
   Take needed number of PCR tube strips out of aluminum bag.
   Important: close bag tightly afterwards. Place strips in a suitable PCR tube rack. If needed, gently tap the tubes to move the lyophilised pellets to the bottom of all tubes.

2. DECAP
   Carefully open strips immediately before filling and discard caps. Do not leave open longer than necessary.

3. ADD SAMPLES AND CONTROLS
   Pipette 5 to 25 µL of samples, 25 µL negative control (colorless cap) or Control Template (purple cap) into respective wells. Varying sample volumes are due to different enrichment broths and DNA extraction procedures. If using less volume, add PCR-grade H2O to reach a total volume of 25 µL.

4. SEAL
   Carefully seal the tubes with the provided 8-cap strips.

5. MIX
   Resuspend pellet after sealing by mixing thoroughly. Alternatively, resuspend pellet by pipetting up and down multiple times in Step 3.

6. CENTRIFUGE
   Briefly spin strips, e.g., 5 seconds at 500 – 1,000 x g, in a suitable centrifuge.

7. 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®
Listeria Genus Detection LyoKit

KIT230088 /089 /330
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

References

• Home | Hygiena

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