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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.
Direct: No RNA Extraction (LDT), straight from VTM/UTM*
Extracted: Use CDC Recommended RNA Extraction
N1 SARS-CoV-2
M1 Influenza A
NS2 Influenza B and human RNaseP genes
5 ul approved VTM/UTM*
or
100 ng to 1 pg extracted RNA (5ul)
N1 SARS-CoV-2 = FAM
M1 FluA = Quasar 670
NS2 FluB = HEX
RNaseP = TexasRed
Amplification 40 cycles,
LoD 5-10 copies
Direct: Heat 100ul VTM 95C for 5’
Indirect: CDC Recommended Protocol
CDC recommended targets and sequences
SARS-CoV-2 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 approximately 1’10”
1. RT Incubation 50C 15’
2. Enzyme Activation 95C 2’
3. Amplification: 40 cycles (95C 3s, 60C 30s)
| SKU# | 20 UL Reactions | Content |
|---|---|---|
| AzureSeq Plus 4-200 | 200 | In Solution: InhibiTaq Plus, Hot Start Mastermix, RTScript, SARS CoV-2 and RNaseP primers and probes, DTT, UDG, RNase Inhibitor |
| AzureSeq Plus 4-1000 | 1000 | |
| AzureSeq Plus 4-2000 | 2000 |
| Target | 20 copies/µL | 10 copies/µL | 5 copies/µL | 1 copy/µL | |
|---|---|---|---|---|---|
| CoV N1 | |||||
| N1 | Positive/Total | 20/20 | 20/20 | 20/20 | 18/20 |
| Ct Average | 33.95 | 35.11 | 36.56 | 39.47 | |
| Influenza A | Positive/Total | 20/20 | 20/20 | 20/20 | 0/0 |
| Ct Average | 34.22 | 35.59 | 37.12 | n/a | |
| Influenza B | Positive/Total | 20/20 | 20/20 | 16/20 | 0/0 |
| Ct Average | 33.01 | 34.43 | 36.58 | n/a |
The inactivated virus was spiked into a negative swab sample plus saline, at 10 copies/uL, 5 copies/uL, and 2.5 copies/uL. Samples were heated at 95C for 5’, then 5uL of sample was added to a final reaction (total 20uL) containing AzureSeq Plus reagents. 20 replicates of each concentration was amplified using recommended cycling conditions (see below). The LOD for the Direct Method (no RNA purification) has been determined to be 10 copies/uL for N1 SARS-CoV-2, M1 Influenza A, and NS2 Influenza B on the BioRad CFX96 Touch Real-time Detection System at least 95% of the time using the AzureSeq Direct Plus RT-qPCR Respiratory Infection Kit with recommended cycling conditions.
| Sample | N1 | FluB | RNaseP | FluA | Result |
|---|---|---|---|---|---|
| CoV-1 | 16.08 | SARS-CoV-2 Detected | |||
| CoV-2 | 23.29 | 25.15 | SARS-CoV-2 Detected | ||
| CoV-3 | 24.96 | 26.51 | SARS-CoV-2 Detected | ||
| CoV-4 | 29.93 | 26.32 | SARS-CoV-2 Detected | ||
| CoV-5 | 27.54 | 30.66 | SARS-CoV-2 Detected | ||
| CoV-6 | 30.12 | 28.95 | SARS-CoV-2 Detected | ||
| CoV-7 | 30.07 | 25.31 | SARS-CoV-2 Detected | ||
| CoV-8 | 22.82 | 27.28 | SARS-CoV-2 Detected | ||
| CoV-9 | 26.26 | 27.73 | SARS-CoV-2 Detected | ||
| CoV-10 | 22.91 | 26.26 | SARS-CoV-2 Detected | ||
| CoV-11 | 16.12 | SARS-CoV-2 Detected | |||
| CoV-12 | 25.19 | 27.08 | SARS-CoV-2 Detected | ||
| CoV-13 | 29.79 | 27.66 | SARS-CoV-2 Detected | ||
| CoV-14 | 21.01 | SARS-CoV-2 Detected | |||
| CoV-15 | 20.19 | 28.65 | SARS-CoV-2 Detected | ||
| CoV-16 | 20.12 | SARS-CoV-2 Detected | |||
| CoV-17 | 18.16 | SARS-CoV-2 Detected | |||
| CoV-18 | 30.26 | 25.12 | SARS-CoV-2 Detected | ||
| CoV-19 | 26.99 | 28.13 | SARS-CoV-2 Detected | ||
| CoV-20 | 21.93 | 31.44 | SARS-CoV-2 Detected | ||
| CoV-21 | 21.36 | SARS-CoV-2 Detected | |||
| CoV-22 | 23.26 | 35.61 | SARS-CoV-2 Detected | ||
| CoV-23 | 38.41 | 27.43 | SARS-CoV-2 Detected | ||
| CoV-24 | 25.65 | 27.31 | SARS-CoV-2 Detected | ||
| CoV-25 | 30.5 | 26.69 | SARS-CoV-2 Detected | ||
| CoV-26 | 23.86 | 43.88 | SARS-CoV-2 Detected | ||
| CoV-27 | 19.78 | 27.3 | SARS-CoV-2 Detected | ||
| CoV-28 | 18.81 | 30.02 | SARS-CoV-2 Detected | ||
| CoV-29 | 30.22 | 30.07 | SARS-CoV-2 Detected | ||
| CoV-30 | 25.46 | 28.73 | SARS-CoV-2 Detected | ||
| CoV-31 | 29.19 | 28.26 | SARS-CoV-2 Detected |
dd
| Target | 20 copies/µL | 10 copies/µL | 5 copies/µL | 1 copy/µL | |
|---|---|---|---|---|---|
| CoV N1 | |||||
| N1 | Positive/Total | 20/20 | 20/20 | 20/20 | 18/20 |
| Ct Average | 33.95 | 35.11 | 36.56 | 39.47 | |
| Influenza A | Positive/Total | 20/20 | 20/20 | 20/20 | 0/0 |
| Ct Average | 34.22 | 35.59 | 37.12 | n/a | |
| Influenza B | Positive/Total | 20/20 | 20/20 | 16/20 | 0/0 |
| Ct Average | 33.01 | 34.43 | 36.58 | n/a |
The inactivated virus was spiked into a negative swab sample plus saline, at 10 copies/uL, 5 copies/uL, and 2.5 copies/uL. Samples were heated at 95C for 5’, then 5uL of sample was added to a final reaction (total 20uL) containing AzureSeq Plus reagents. 20 replicates of each concentration was amplified using recommended cycling conditions (see below). The LOD for the Direct Method (no RNA purification) has been determined to be 10 copies/uL for N1 SARS-CoV-2, M1 Influenza A, and NS2 Influenza B on the BioRad CFX96 Touch Real-time Detection System at least 95% of the time using the AzureSeq Direct Plus RT-qPCR Respiratory Infection Kit with recommended cycling conditions.
| Sample | N1 | FluB | RNaseP | FluA | Result |
|---|---|---|---|---|---|
| CoV-1 | 16.08 | SARS-CoV-2 Detected | |||
| CoV-2 | 23.29 | 25.15 | SARS-CoV-2 Detected | ||
| CoV-3 | 24.96 | 26.51 | SARS-CoV-2 Detected | ||
| CoV-4 | 29.93 | 26.32 | SARS-CoV-2 Detected | ||
| CoV-5 | 27.54 | 30.66 | SARS-CoV-2 Detected | ||
| CoV-6 | 30.12 | 28.95 | SARS-CoV-2 Detected | ||
| CoV-7 | 30.07 | 25.31 | SARS-CoV-2 Detected | ||
| CoV-8 | 22.82 | 27.28 | SARS-CoV-2 Detected | ||
| CoV-9 | 26.26 | 27.73 | SARS-CoV-2 Detected | ||
| CoV-10 | 22.91 | 26.26 | SARS-CoV-2 Detected | ||
| CoV-11 | 16.12 | SARS-CoV-2 Detected | |||
| CoV-12 | 25.19 | 27.08 | SARS-CoV-2 Detected | ||
| CoV-13 | 29.79 | 27.66 | SARS-CoV-2 Detected | ||
| CoV-14 | 21.01 | SARS-CoV-2 Detected | |||
| CoV-15 | 20.19 | 28.65 | SARS-CoV-2 Detected | ||
| CoV-16 | 20.12 | SARS-CoV-2 Detected | |||
| CoV-17 | 18.16 | SARS-CoV-2 Detected | |||
| CoV-18 | 30.26 | 25.12 | SARS-CoV-2 Detected | ||
| CoV-19 | 26.99 | 28.13 | SARS-CoV-2 Detected | ||
| CoV-20 | 21.93 | 31.44 | SARS-CoV-2 Detected | ||
| CoV-21 | 21.36 | SARS-CoV-2 Detected | |||
| CoV-22 | 23.26 | 35.61 | SARS-CoV-2 Detected | ||
| CoV-23 | 38.41 | 27.43 | SARS-CoV-2 Detected | ||
| CoV-24 | 25.65 | 27.31 | SARS-CoV-2 Detected | ||
| CoV-25 | 30.5 | 26.69 | SARS-CoV-2 Detected | ||
| CoV-26 | 23.86 | 43.88 | SARS-CoV-2 Detected | ||
| CoV-27 | 19.78 | 27.3 | SARS-CoV-2 Detected | ||
| CoV-28 | 18.81 | 30.02 | SARS-CoV-2 Detected | ||
| CoV-29 | 30.22 | 30.07 | SARS-CoV-2 Detected | ||
| CoV-30 | 25.46 | 28.73 | SARS-CoV-2 Detected | ||
| CoV-31 | 29.19 | 28.26 | SARS-CoV-2 Detected |
dd
Suggested CPT Code: 87636 Infectious agent detection by nucleic acid (DNA or RNA); severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) (Coronavirus disease [COVID-19]) and influenza virus types A and B, multiplex amplified probe technique.
Suggested CPT Code: 87636 Infectious agent detection by nucleic acid (DNA or RNA); severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) (Coronavirus disease [COVID-19]) and influenza virus types A and B, multiplex amplified probe technique.
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.
It is available for custom synthesis
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
For validation N1=5 copies/rxn, N2=10 copies/rxn.
1000 cp/ul RNaseP, 200 cp/ul of N
| 2019-nCoV_N1-P | 2019-nCoV_N1 Probe | FAM-ACC CCG CAT TAC GTT TGG TGG ACC-BHQ1 | FAM, BHQ-1 | 125nM |
| 2019-nCoV_N2-P | 2019-nCoV_N2 Probe | FAM-ACA ATT TGC CCC CAG CGC TTC AG-BHQ1 | FAM, BHQ-1 | 125nM |
| RP-P | RNAse P Probe | FAM – TTC TGA CCT GAA GGC TCT GCG CG – BHQ-1 | FAM, BHQ-1 | 125nM |
https://www.cdc.gov/coronavirus/2019-ncov/lab/rt-pcr-panel-primer-probes.html
60mer-70mer
N1, N2 capsid and RP-P
It was validated using standard qPCR instruments including: QuantStudio 7 Flex series 96/384, BioRad CFX96/384 and ABI 7500 Fast Dx.
