AZURESEQ
ONE STEP UNIVERSAL RT-qPCR Kit
SARS-CoV-2
AZURESEQ PLUS
ONE STEP UNIVERSAL RT-qPCR Kit
SARS-CoV-2, INFLUENZA A/B
Get high multiplex performance in an American-made product. Excellent sensitivity with CDC-recommended probe/primer sequences. All enzymes are synthesized in the United States.
Two Protocols
Direct: No RNA Extraction (LDT)
Indirect: Use CDC Recommended Extraction
Choose your format – Direct or RNA Extraction
Increase your throughput 25% to 300%
Fewer steps reduce potential user error
Save on extraction costs
Uses common qPCR channels
Supply chain continuity with American products
AzureSeq
Targets/Dye
N1 SARS-CoV-2=FAM
N2 SARS-CoV-2= HEX
RNaseP=TxRed
Sample
5ul of approved VTM/UTM*
or
100ng to 1pg RNA from
CDC recommended protocol
AzureSeq Plus 4
Targets/Dye
N1 SARS-CoV-2=FAM
M1 Influenza A= Quasar 670
NS2 Influenza B= HEX
RNaseP=TxRed
* Approved VTM/UTM: Copan, Puritan, CDC, Saline, PBS, MAWI EL
AzureSeq vs. AzureSeq Plus 4
| Pathogens | Dyes | Sample Processing | |
|---|---|---|---|
| AzureSeq | SARS-CoV-2 | HEX, FAM, TxRed | Direct & CDC RNA Ext Protocols |
| AzureSeq Plus 4 | SARS-CoV-2, Influenza A/B | HEX, FAM, ROX, Quasar 670 | Direct & CDC RNA Ext Protocols |
Indirect: CDC Recommended Protocol
CDC recommended targets and sequences
Use Cy5 channel to detect Quasar 670
HEX, ROX, FAM channels standard
QuantStudio Series, BioRad CFX, Roche Lightcycler
5-10 copies per reaction using CDC RNA extraction protocol, 10 copies per reaction using direct protocol.
Influenza A/B
25 copies per reaction using Direct Protocol
40 cycles using default ramp rates
Total cycling time appx 1’10”
- RT Incubation 50C 15’
- Enzyme Activation 95C 2’
- Amplification: 40 cycles (95C 3s, 60C 30s)
Reaction sizes: 200, 1000, 2000, custom
PERFORMANCE EVALUATION
Contrived Clinical Samples
Competitor Comparison
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.
AzureSeq Plus Simulated Co-Infection Study
AzureSeq LoD Data
| 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.
| 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 (20 replicates each conc) 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.
AzureSeq
- Instructions for Use – With Purification
- Instructions for Use – Direct
- Instructions for Use – Validation Kit
- Health Care Provider Fact Sheet
- Patient Fact Sheet
AzureSeq Plus 4
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.