AZURESEQ
ONE STEP UNIVERSAL RT-qPCR Kit
SARS-CoV-2
Get high multiplex performance in an American-made product. Excellent sensitivity with CDC-recommended primers. All enzymes are synthesized in the United States. Fast turn-around times for customization that fits your workflow and format.
Detect SARS-CoV-2
in Less Than One Hour
Single-Tube RT-qPCR Detection
Made in America
Research Use Only
Accurate detection with reduced complexity, time and waste.
Low copy number (5-10 copies) delivers a reliable, highly sensitive one-step RT-qPCR solution for rapid testing.
Compatible with CDC recommended RNA extraction kits and qPCR instruments.
AzureSeq
Multiplex Detection of
SARS-CoV-2
This multiplex RT-qPCR assay detects two regions on the SARS-CoV-2 nucleocapsid (N) gene, N1 and N2, on two different channels. The assay uses human RNase P as the sample control gene on the third channel
Two Protocols – Same Kit
Direct: No RNA Extraction (LDT), straight from VTM/UTM*
Extracted: Use CDC Recommended RNA Extraction
Targets
2 regions of SARS-CoV-2 virus N nucleocapsid protein gene plus control region of RNaseP gene
Sample
100ng to 1pg using CDC recommended extraction protocols
Dyes/Cycles
N1=FAM, N2=HEX, RNaseP=TxRed/ROX, 40 Amplification cycles,
LoD: 5-10 copies
Parameters
Amplification 40 cycles,
LoD 5-10 copies
*Approved VTM/UTM: Copan, Puritan, CDC, Saline, PBS, MAWI EL
PERFORMANCE EVALUATION
Contrived Clinical Samples
AzureSeq was determined to be compatible with RNA extraction workflow at concentrations at or near LoD. Pooled negative OP swabs were spiked with genomic RNA. Input volume for extraction was 250 μl and elution volume was 80 μl. A 5 μl aliquot of the eluate was added to the PCR reaction.
-
- nCoV2 N1
-
- nCoV2 N2
Detection on nCoV2 N1 and N2 genes: 50 copies of genomic RNA added in a 20 μl reaction. Reaction set-up and RT-qPCR protocol were in accordance with CDC guidance.
| N1 | N2 | Rnase P | |||||||
|---|---|---|---|---|---|---|---|---|---|
| Copies | Cp | Average | Std Dev | Cp | Average | Std Dev | Cp | Average | Std Dev |
| 40 cp/uL | 33.29 | 33.38 | 0.11 | 33.97 | 34.08 | 0.10 | 32.92 | 32.47 | 0.48 |
| 40 cp/uL | 33.34 | 34.13 | 31.96 | ||||||
| 40 cp/uL | 33.50 | 34.14 | 32.52 | ||||||
| 20 cp/uL | 34.04 | 34.27 | 0.20 | 34.81 | 35.13 | 0.29 | 31.97 | 31.96 | 0.22 |
| 20 cp/uL | 34.41 | 35.24 | 31.73 | ||||||
| 20 cp/uL | 34.35 | 35.35 | 32.17 | ||||||
| 10 cp/uL | 34.51 | 34.87 | 0.36 | 35.63 | 36.03 | 0.37 | 32.13 | 32.04 | 0.11 |
| 10 cp/uL | 34.88 | 36.35 | 32.07 | ||||||
| 10 cp/uL | 35.22 | 36.12 | 31.92 | ||||||
| 5 cp/uL | 37.01 | 36.86 | 0.70 | 37.37 | 37.88 | 1.10 | 32.72 | 32.39 | 0.33 |
| 5 cp/uL | 36.10 | 37.12 | 32.06 | ||||||
| 5 cp/uL | 37.48 | 39.14 | 32.40 | ||||||
| 1 cp/uL | 37.67 | 38.32 | 0.89 | 40.38 | 39.27 | 0.99 | 32.39 | 32.34 | 0.10 |
| 1 cp/uL | 37.96 | 38.48 | 32.41 | ||||||
| 1 cp/uL | 39.33 | 38.95 | 32.23 |
Legend
Negative nasopharyngeal swabs were spiked with heat-killed SARS-CoV-2 virus at the labeled concentration in copies/uL in starting sample media (saline). Samples were heated at 95C for 5 min, then 5uL of heat-inactivated sample was added to reactions containing 1X AzureSeq CoV-2 reagents. The final reaction volumes were 20ul. Samples were run on a BioRad CFX96 using default ramp rates.
The LoD for the AzureSeq One-Step reagents for genomic RNA on the BioRad CFX96 Touch Real-time Detection System has been determined to be 5 copies per 20-μL reaction (or 0.25 copies/μL) for N1 and 10 copies per 20-μL reaction (or 0.5 copies/μL) for N2. Genomic RNA was spiked directly into PCR reactions and then analyzed on the BioRad CFX96 Touch Real- Time Detection System.
The LoD for the AzureSeq One-Step reagents for genomic RNA on the Applied Biosystems QuantStudio 3 Real-time PCR System has been determined to be 10 copies per 20-μL reaction (or 0.5 copies/μL) for both N1 and N2 targets. Genomic RNA was spiked directly into PCR reactions and then analyzed on the Quant-Studio Real-Time PCR System.
ORDERING INFORMATION
| SKU# | 20 ul Reactions | Content |
|---|---|---|
| AzureSeq Direct-200 | 200 | In Solution: InhibiTaq Plus, Hot Start Mastermix, RTScript, SARS CoV-2 and RNaseP primers and probes, DTT, UDG, RNase Inhibitor |
| AzureSeq Direct-1000 | 1000 | |
| AzureSeq Direct-2000 | 2000 | |
| AzureSeq Direct Custom | Custom |
For Research Use Only. CLIA labs must file an LDT or EUA.
Depending on your cycler, approximately 70′-75′.
For N1, FAM (absorption 493nm, emission 517nm)
For N2, HEX (absorption 533nm, emission 559nm)
For RnaseP, ROX/Tx Red (absorption 583nm, emission 603nm)
Yes. Our FDA Facility Registration number is 10076042
Yes, we can make AzureSeq in tube and 96/384-well format.
- We anticipate mid-August 2020. Samples are available now for testing.
- Will you submit for EUA? Yes, assuming validation data is comparable or superior to the wet version. It is currently RUO- Not For Diagnostic Use.
As of Jul 12, 2020 we can provide 2M reactions/week. This can be further scaled with 10 days notice. Please contact us for the most up-to-date information.
Both are available.
Omega Total RNA Spin Columns and KingFisher MagMax
For validation N1=5 copies/rxn, N2=10 copies/rxn.
1000 cp/ul RNaseP, 200 cp/ul of N
60mer-70mer
N1, N2 capsid and RP-P
It was validated using standard qPCR instruments including: QuantStudio series, BioRad FX.
It is EUA-Validated, not EUA-Authorized. At this time, CLIA users may choose to submit their own EUA. Upon request, we will send the FDA EUA Submission for AzureSeq.
Product Disclaimer
Download SDS Sheets
- Safety Data Sheet – Nuclease Free Water
- Safety Data Sheet – CoVi Positive Control
- Safety Data Sheet – CoVi Primer/Probe Mix 3
- Safety Data Sheet – RTScript, 200U/uL/WarmStart RTScript, 200U/uL
- Safety Data Sheet – 2X InhibiTAQ Multiplex HotStart MasterMix
- Safety Data Sheet – CoVi Negative Control
- Safety Data Sheet – Direct RT Mix