hygiena KIT230043 Enterobacteriaceae Plus Cronobacter Detection Kit User Guide
- May 15, 2024
- Hygiena
Table of Contents
foodproof®
Enterobacteriaceae plus Cronobacter Detection Kit
Ready Reference Guide
Product No. KIT230043
PCR kit for the qualitative detection of Enterobacteriaceae plus simultaneous
identification of Cronobacter spp.
DNA 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 (Cronobacter), HEX (Enterobacteriaceae) and ROX (Internal Control).
Pre-incubation: 1 cycle
Step 1: 37 °C for 4 min
Step 2: 95 °C for 5 min
Amplifi cation: 40 cycles
Step 1 : 95 °C for 10 sec
Step 2*: 65 °C for 70 sec,
Step down each cycle by 0.1°C
- Fluorescence detection
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 results of positive (Control Template) and negative (H2 O) 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.
| FAM | HEX | ROX | Result Interpretation |
|---|---|---|---|
| + | + | + or – | Positive for Cronobacter and Enterobacteriaceae |
| – | + | + or – | Positive for Enterobacteriaceae |
| – | – | + | Negative for Cronobacter and Enterobacteriaceae |
| – | – | – | Invalid |
PREPARATION OF THE PCR MIX
Take appropriate precautions to prevent contamination, e.g., by using fi lter
tips and wearing gloves.
Thaw reagents, mix (do not vortex!) and briefl y spin vials before opening.
-
PREPARE PCR MIX
Add 18 µL of Master Mix (yellow cap), 1 µL of Enzyme Solution (red cap) and 1 µL of Internal Control (white 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 PCR MIX
Pipet 20 µL of prepared PCR mix into each strip or plate well. -
ADD SAMPLES AND CONTROLS
Pipet 5 µL of samples, Negative Control (colorless 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.
foodproof®
Enterobacteriaceae plus
Cronobacter Detection Kit
KIT230043
Kit for 96 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