AZURESEQ & AZURESEQ PLUS

ONE STEP UNIVERSAL RT-qPCR KITS
LYOPHILIZED FORMATS

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.

LYOPHILIZED FORMATS

Direct: No RNA Extraction
Indirect: Use CDC Recommended Extraction

RUO, Not For Diagnostics
CLIA Labs Must File LDT or EUA

  • 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
N2 SARS-CoV-2=HEX
M1 Influenza A= Quasar 670
NS2 Influenza B= Quasar 670
RNaseP=TxRed

Lyophilized vs. Wet Comparison

 SpeedEaseTemperature
Lyophilized FormatFastest: Less than 90 minEasiest: Resuspend - GoRoom Temp: Stable at 37ºC
Wet FormatFast: Less than 120 minEasy: 8 pipetting stepsFrozen: Store at -20ºC

AzureSeq vs. AzureSeq Plus 4

 PathogensDyesSample Processing
AzureSeqSARS-CoV-2HEX, FAM, TxRedDirect & CDC RNA Ext Protocols
AzureSeq Plus 4SARS-CoV-2,
Influenza A/B
HEX, FAM, ROX, Quasar 670Direct & CDC RNA Ext Protocols

Important
AzureSeq Plus does not differentiate between Influenza A and Influenza B.

Direct: Heat 100ul VTM 95C for 5’
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

SARS-CoV-2
5-10 copies per reaction using CDC RNA extraction protocol

40 cycles using default ramp rates
Total cycling time appx 1’45”

  1. RT Incubation 50C 15’
  2. Enzyme Activation 95C 2’
  3. Amplification: 40 cycles (95C 3s, 60C 30s)

Available in user-friendly, custom formats.

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.

Wet vs. Lyophilized

AzureSeq (wet) and AzureSeq Lyo (lyophilized) show comparable results using CDC specified primers and probes for nCoV2 N1 and N2 and RPP controls using CDC recommended testing protocol.

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 LoD Data

 N1  N2  Rnase P  
CopiesCpAverageStd DevCpAverageStd DevCpAverageStd Dev
40 cp/uL33.2933.380.1133.9734.080.1032.9232.470.48
40 cp/uL33.3434.1331.96
40 cp/uL33.5034.1432.52
20 cp/uL34.0434.270.2034.8135.130.2931.9731.960.22
20 cp/uL34.4135.2431.73
20 cp/uL34.3535.3532.17
10 cp/uL34.5134.870.3635.6336.030.3732.1332.040.11
10 cp/uL34.8836.3532.07
10 cp/uL35.2236.1231.92
5 cp/uL37.0136.860.7037.3737.881.1032.7232.390.33
5 cp/uL36.1037.1232.06
5 cp/uL37.4839.1432.40
1 cp/uL37.6738.320.8940.3839.270.9932.3932.340.10
1 cp/uL37.9638.4832.41
1 cp/uL39.3338.9532.23

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 
CopiesCpStd DevCpStd DevCpStd Dev
20 cp/uL34.050.2034.910.2832.060.22
10 cp/uL34.910.3036.020.3532.080.21
5 cp/uL36.530.6237.580.6032.530.35

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.

ORDERING INFORMATION

AzureSeq Lyo and AzureSeq Plus Lyo is for RUO only. CLIA labs must file an LDT or EUA.

AzureSeq

AzureSeq Plus 4

  • Instructions for Use – With Purification
  • Instructions for Use – Direct
  • Instructions for Use – Validation Kit
  • Health Care Provider Fact Sheet
  • Patient Fact Sheet

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

DOING OUR PART

By highlighting threatened or endangered species, SeqOnce hopes to draw attention to conservation efforts. As biologists, we believe humanity is better served when ecologies are maintained and there is diversity of life.

SeqOnce supports protecting and conserving endangered and threatened species.

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