GENERATION Biotech VoXcreen Delta Omicron SARS-CoV-2 Screening and Variant Detection Kit Instructions
- June 13, 2024
- GENERATION Biotech
Table of Contents
VoXcreen Delta / Omicron
SARS-CoV-2 Screening & Variant Detection Kit
Instructions for Use
Application
The “VoXcreen” SARS-CoV-2 Delta / Omicron Screening & Variant Detection Kit is
a reagent system, based on quantitative reverse transcriptase (qRT) PCR
technology, for the detection of RNA specific to SARS-CoV2 and its Variants of
Concern (VoC) that cause the Coronavirus Disease 2019 (COVID-19).
VoXcreen utilizes a set of variant-specific viral mutations to detect and
discriminate in a single step the most important current Variants of Concern
(VoC): Delta and Omicron. For the detection of variants other than Delta or
Omicron we recommend to use our follow-up assays.
Quantitative polymerase chain reaction (qPCR) technology utilizes an enzyme
(reverse-transcriptase, RT) to convert RNA to complementary DNA (cDNA), from
which variant-specific target sequences are then amplified and targeted with
specific probes for the detection of their copy number in the initial
specimen.
For Research Use Only. Not for use in diagnostic procedures. Not FDA approved
or CE certified.
Equipment and Consumables
Customer supplied equipment
Real-Time PCR Cycler
Calibrated single-channel pipettes
Vortex mixer
Benchtop microcentrifuge for 1.5 ml tubes
Customer supplied consumables
Pipette tips (with aerosol barriers, nuclease-free)| General laboratory
supplier
---|---
Real-Time PCR Plates| General laboratory supplier
1.5 ml tubes (low DNA binding, nuclease-free)| General laboratory supplier
Kit Components
| Component | Tube Label |
|---|---|
| Buffer Mix | BM |
| Enzyme Mix | EM |
| Primer / Probe Mix | PPM |
| Positive Control WT | CTR WT |
| Positive Control Delta | CTR Delta |
| Positive Control Omicron | CTR Omicron |
| Nuclease-free water | Water |
Storage
SARS-CoV-2 Screening & Variant Detection Kits are shipped on -20° C or -80°C
ice packs. The components of the kit should arrive frozen. If one or more
components are not frozen upon receipt, or if tubes have been compromised
during shipment, contact Generation Biotech for assistance.
All components should be stored between -25° C and -15° C upon arrival.
Repeated thawing and freezing of reagents should be avoided, as this might
affect the performance of the assay. The reagents should be subdivided and
frozen in aliquots if they are to be used intermittently.
Storage between +2° C and +8° C should not exceed a period of two hours.
Protect Primer/Probe Mixes (PPM, blue cap) from light.
Product Description
The SARS-CoV-2 Delta / Omicron Variant Detection Assay determines the presence
of the Delta or Omicron strains of SARS-CoV-2 by discriminating between
variant-specific deletions.
The SARS-CoV-2 Delta / Omicron Variant Detection Assay consists of:
| Buffer Mix | Mix of buffer and dNTPs |
|---|---|
| Master Mix | Mix of reverse transcriptase and heat-stable polymerase |
| Primer / Probe Mix | Mix of primers and probes |
| Positive Controls | SARS-CoV-2 RNA of WT, Delta and Omicron with human RNase P |
| Diluent | Nuclease-free Water |
The assay consists of three processes in a single reaction tube (one-step qRT- PCR):
- Reverse transcription of target RNA (SARS-CoV-2 and human RNase P) to cDNA
- PCR amplification of target cDNA
- Quantitative detection of PCR amplification products by fluorescent dye labeled probes FAM, HEX, ROX and CY5.
Procedure
6.1 Sample Preparation
RNA extracted from the specimen is the starting material for the SARS-CoV-2
Variant Detection Kit.
The quality of the extracted RNA has a profound impact on the performance of
the entire test system.
It is recommended to ensure that the system used for nucleic acid (RNA/DNA)
extraction is compatible with the characteristics of the sample (i.e. the
presence of contaminants, such as in wastewater) and with the desired type of
quantitative PCR technology and instrumentation.
If using a spin column-based sample preparation procedure that includes
washing buffers containing ethanol, it is highly recommended to perform an
additional centrifugation step for 5 min. at maximum speed, using a new
collection tube and prior to the elution of the nucleic acid, to obtain
reliable results.
For additional information and technical support regarding pre-treatment and
sample preparation please contact our Technical Support (see Technical
Assistance).
6.2 Reaction Setup
All reagents and samples should be thawed completely, mixed (by pipetting or
gentle vortexing) and centrifuged briefly before use.
| Reagents | Volume per Reaction |
|---|---|
| Buffer Mix (BM) | 10 µl |
| Enzyme Mix (EM) | 0.2 µl |
| Primer / Probe Mix (PPM) | 2 µl |
| Sample or Positive Control (CTR) | 5 µl |
| Nuclease-free Water | 2.8 µl |
| Total Reaction Volume | 20 µl |
Programming the Real-Time PCR Instrument
For basic information regarding the setup and programming of the different
real-time PCR instruments, please refer to the user manual of the respective
instrument.
7.1 Settings
Define the following settings as described below.
Settings
| Reaction Volume | 20 µl |
|---|---|
| Ramp Rate | Default |
| Passive Reference | (None) |
7.2 Fluorescence Detector
Define the fluorescence detectors as described below.
| Target | Reporter Dye | Quencher Dye |
|---|---|---|
| SARS-CoV-2 gene N1 | FAM | (none) |
| Human RNase P | HEX | (none) |
| Human RNase P | ROX | (none) |
| SARS-CoV-2 Delta specific deletion | CY5 | (none) |
7.3 Temperature Profile
Incubate the reaction as described below.
| Step | Stage | Cycles | Acquisition | Temp | Time |
|---|---|---|---|---|---|
| RT | Hold | 1 | – | 50 °C | 10 min |
| Denaturation | Hold | 1 | – | 95 °C | 2 min |
| Denaturation | Cycling | 50 | – | 95 °C | 5 sec |
| Amplification | YES | 60 °C | 30 sec |
Data Analysis
For basic information regarding data analysis on specific real-time PCR
instruments, please refer to the user manual of the respective instrument. For
detailed instructions regarding the analysis of the data generated with the
SARS-CoV-2 Variant Detection Kit on different quantitative PCR instruments
please contact our Technical Support. Please adjust the threshold of your qPCR
instrument for positive detection to be at 10% of the signal intensity of the
maximum value of the positive control. Samples that reach less
than 10% of the signal intensity of the positive control shall be regarded as
negative.
8.1 Interpretation of
Results
CoV-2 N1 FAM| Human RNase P HEX| CoV-2 Omicron ROX| CoV-2
Delta CY5| Result
---|---|---|---|---
POS| POS or NEG| POS| NEG| SARS-CoV-2
Omicron Variant Detected
POS| POS or NEG| NEG| POS| SARS-CoV-2
Delta Variant Detected
POS| POS or NEG| NEG| NEG| Neither Omicron
or Delta Variant Detected
NEG| POS| NEG| NEG| SARS-CoV-2
Not Detected
NEG| NEG| NEG| NEG| Invalid
(Repeat Test)
Technical Assistance
For customer support, please contact our Technical Support: e-mail:
[email protected]
phone: 609-637-0878
Disclaimers
For Research Use Only. Not for use in diagnostic procedures. Not FDA approved
or CE certified.
Registered names, trademarks, etc. used in this document, even if not
specifically marked as such, are not to be considered unprotected by law.
VoXcreen SARS-CoV-2 Screening & Variant Detection Kit
Instructions for Use – 20211231
Generation Biotech
31 Airpark Road
Princeton, NJ 08540
www.gen-bio.com