hygiena KIT230145 LP Aspergillus Detection LyoKit User Guide
- May 15, 2024
- Hygiena
Table of Contents
hygiena KIT230145 LP Aspergillus Detection LyoKit
Specifications
- Product Name: Aspergillus Detection LyoKit
- Revision: A, February 2024
- Product Numbers: KIT230145 (LP), KIT230146 (RP), KIT230147 (DP)
- PCR kit for the qualitative detection of Aspergillus flavus, A. terreus, A. niger, and A. fumigatus
Program Setup
Program your real-time PCR instrument before setting up the PCR reactions.
Select the following channels:
- FAM (A. flavus)
- HEX (A. terreus)
- ROX (A. niger and A. fumigatus)
- Cy5 (Internal Control)
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 provided in the manual.
Preparation of the PCR Mix
- PLACE STRIPS IN RACK : Take needed number of PCR tube strips out of the aluminum bag. Place strips in a suitable PCR tube rack.
- DECAP : Carefully open strips immediately before filling and discard caps.
- ADD SAMPLES AND CONTROLS : Add the sample into wells following the detailed protocols in the Product Instructions.
- SEAL : Carefully seal the tubes with the provided 8-cap strips.
- MIX : Resuspend pellet after sealing by mixing thoroughly or by pipetting up and down multiple times.
- CENTRIFUGE : Briefly spin strips in a suitable centrifuge.
- START REAL-TIME PCR RUN : Cycle samples as described above. Place tubes in a vertical, balanced order into the cycler.
FAQ
-
Q: What should I do if my sample results are inconclusive?
A: If your sample results are inconclusive, it is recommended to repeat the test following the provided protocols carefully to ensure accurate results. -
Q: Can I reuse the PCR tube strips?
A: It is not recommended to reuse PCR tube strips to avoid any potential contamination issues. Always use fresh strips for each test. -
Q: How should I store the LyoKit when not in use?
A: Store the LyoKit in a cool, dry place away from direct sunlight. Ensure that the kit is properly sealed and protected from moisture.
foodproof®
Aspergillus Detection LyoKit
Revision A, February 2024
Ready Reference Guide
Product No. KIT230145 (LP), KIT230146 (RP), KIT230147 (DP)
PCR kit for the qualitative detection of Aspergillus flavus, A. terreus, A.
niger and A. fumigatus.
Before starting, it is strongly recommended to read the entire product manual
available on our website. For the BAX® Q7 in combination with the BAX Prep
Aspergillus Lysis Kit (KIT2046), please use the separately available
instructions.
PROGRAM SETUP
Program your real-time PCR instrument before setting up the PCR reactions.
Select the following channels
FAM (A. flavus), HEX (A. terreus), ROX (A. niger and A. fumigatus) and Cy5 (Internal Control).
- Pre-incubation: 1 cycle Step 1: 37 °C for 4 min Step 2: 95 °C for 5 min
- Amplifi cation: 50 cycles Step 1: 95 °C for 5 sec Step 2*: 60 °C for 60 sec Step 3: 72 °C for 30 sec
- Melting Curve : 1 cycle Step 1: 95 °C for 50 sec Step 2: 37 °C for 50 sec Step 3**: ramp up to 85 °C
- Fluorescence detection
- Fluorescence detection during 37 to 85 °C ramp with 1 to 2 measurements/°C
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. Color Compensation Set 5 (Product No. KIT230011) is necessary for users of the LightCycler® 480 System. 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.
Amplification Curve| FAM| HEX| ROX| Cy5| Result
Interpretation
---|---|---|---|---|---
+| –| –| +/-| Positive for Aspergillus flavus
–| +| –| +/-| Positive for Aspergillus terreus
–| –| +| +/-| Positive for Aspergillus niger and/or A. fumigatus
–| –| –| +| Negative for all tested Aspergillus species
–| –| –| –| Invalid
Differentiation
ng e| Channel| Aspergillus niger| Aspergillus
fumigatus
---|---|---|---
Melti Curv| ROX| 62 ± 3 °C| **** 73 ± 3 °C
PREPARATION OF THE PCR MIX
Take appropriate precautions to prevent contamination, e.g., by using filter tips and wearing gloves. Please refer to the Product Instructions for each kit for additional information.
- 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 lyophilized pellets to the bottom of all tubes.
- DECAP
- Carefully open strips immediately before filling and discard caps.
- Do not leave open longer than necessary.
- ADD SAMPLES AND CONTROLS
- Pipette 25 µL of negative control (colorless cap) or Control Template (purple cap) into respective wells.
- For extractions using StarPrep® Two Kit: Pipette 20 µL PCR-grade H2O and 5 µL sample into wells.
- For extractions using BAX Prep Aspergillus Lysis Kit: Pipette 30 µL of sample into wells.
- (Please see Product Instructions for the respective kits for detailed protocols).
- SEAL
- Carefully seal the tubes with the provided 8-cap strips.
- MIX
- Resuspend pellet after sealing by mixing thoroughly.
- Alternatively, resuspend pellet by pipetting up and down multiple times in Step 3.
- CENTRIFUGE
- Briefl y 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® Aspergillus Detection LyoKit
KIT230145 /46 /47 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