COVID-19 Diagnostic Workflow
Collection & Preservation
Viral RNA Isolation
PCR Based Detection
PCR Components
1. SAMPLE COLLECTION & PRESERVATION
Total Nucleic Acid Preservation Tubes Dx
- CE-IVD marked in accordance with EU Directive 98/79/EC
- Ideal for use in in vitro diagnostic workflows
- No need to immediately process samples
- Total DNA, including viral DNA, preserved at room temperature over 4 months
- Total RNA, including viral RNA, preserved at room temperature over 2 months
- Inactivate microorganisms including bacteria, fungi, yeast and viruses
- Ship swab samples at room temperature safely
- Compatible with most DNA and Viral RNA isolation methods
Preserve High Quality RNA for over 2 Months and DNA for over 4 Months at Ambient Temperature
Figure 1. Stability of RNA and DNA stored in Norgen’s Total Nucleic Acid Preservative. GAPDH (for DNA) and β-actin (for RNA) were monitored over an 18 week period for DNA and 60 days for RNA from swab samples stored at ambient temperature in Norgen’s Total Nucleic Acid Preservation Tubes. RNA was isolated from 250 µL of preserved swab samples on day 0, day 4, day 6, day 10, day 16, day 30 and day 60 using Norgen’s Total RNA Isolation Kit (Cat.# 17200). DNA was isolated from 250 µL of preserved swab samples on day 0 and 18 weeks using Norgen’s Saliva DNA Isolation Kit (Cat# Dx45400). Next, 4 µL of elution was used in a 20 µL real-time PCR reaction using CFX96 Touch™ Real-Time PCR Detection System (Bio-Rad). RNA can be stored in Norgen’s Total Nucleic Acid Preservative for up to 60 days and DNA can be stored for up to 18 weeks at room temperature with no change in quality and quantity of RNA and DNA.
| Cat. # | Description | Size | |
|---|---|---|---|
| Cat. Dx69200 | Total Nucleic Acid Preservation Tubes Dx (For Diagnostic Use) | 50 units | View product page |
Option 2: Saliva RNA Collection and Preservation Devices Dx
| Cat. # | Description | Size | |
|---|---|---|---|
| Cat. 53800 | Saliva RNA Collection and Preservation Devices Dx (For Diagnostic Use) | 50 devices | View product page |
Proceed to Viral RNA isolation
2. VIRAL RNA ISOLATION
Saliva/Swab RNA Purification Kits Dx
- CE-IVD marked in accordance with EU Directive 98/79/EC
- Fits into in vitro diagnostic workflows
- Isolate high quality total RNA, including viral RNA, from fresh and preserved saliva and swab samples
- Fast and easy processing using rapid spin-column format
- Isolate total RNA, from large rRNA down to microRNA (miRNA)
- No phenol or chloroform extractions
- Very sensitive & linear down to a few cells/viral copies without the need for carrier RNA
- High throughput 96-well plate format for rapid purifications is also available
- Purified RNA is suitable for a variety of downstream applications, including Small RNA Sequencing. Find out more information on Norgen’s NGS services
Isolate High Quality Total RNA, including Viral RNA, from Fresh & Preserved Saliva and Swab Samples
Figure 1. SARS-CoV-2 E-gene and RdRP Amplification from Preserved Swabs and Saliva RNA isolated by Norgen’s Saliva/Swab RNA Purification Kit (Cat. #69100). Duplicate Nasopharyngeal swabs, Oropharyngeal Swabs and Saliva samples were collected from different donors. Nasopharyngeal and Oropharyngeal swab samples were collected into Norgen’s Total Nucleic Acid Preservative Tubes (Cat. 69200). Saliva samples were collected into Norgen’s Saliva RNA Collection and Preservation Devices (Cat. RU53800). All collected samples were spiked with 3 different concentrations from Norgen’s E/RdRP/RP Positive Control (included in Norgen’s COVID-19 TaqMan RT-PCR Kit (E/RdRP genes – TM67200) and processed for RNA isolation using Norgen’s Saliva/Swab RNA Purification Kit (Cat. #69100), using an input of 0.25 mL from each preserved samples. The sensitivity of the isolation method was assessed amplifying the spiked SARS-CoV-2 spiked targets using Norgen’s COVID-19 TaqMan RT-PCR Kit (E/RdRP genes (TM67200). Figure 1 shows the linear recovery of the different spike-in amounts indicating the high sensitivity of the isolation procedure as well as the high quality of the RNA purified using Norgen’s Saliva/Swab RNA Purification Kit (Cat. #69100).
| Cat. # | Description | Size | |
|---|---|---|---|
| Dx69100 | Saliva / Swab RNA Purification Kit Dx | 50 Preps | View product page |
| Cat. Dx69300 | Saliva/Swab RNA Purification 96-Well Kit Dx | 2 x 96-well plates | View product page |
Option 2: Total Purification Kits Dx
| Cat. # | Description | Size | |
|---|---|---|---|
| Dx17200 | Total RNA Purification Kit Dx | 50 Preps | View product page |
| 17200, 37500, 17250, 17270 |
Total RNA Purification Kit | 50 Preps, 100 preps, 250 preps, 500 preps |
View product page |
| 24300, 24370 24350, 24380 |
Total RNA Purification 96-Well Kits | 2 x 96-well plates, 6 x 96-well plates |
View product page |
3. PCR BASED DETECTION
COVID-19 TaqMan RT-PCR Kit (E/RdRP genes)
- SARS-CoV-2 detection based on the Charité/Berlin protocol (E/RdRP genes) recommended by WHO.
- Primer & Probe Mixes contain all E gene/RP and RdRP gene assays in individual tubes
- All assays are premixed to the working concentrations recommended by the Charité/Berlin protocol.
- Positive Control contains the three SARS-related target genes: E gene, RdRP gene as well as the human RP (internal control)
- Convenient ready-to-use 2X One-Step RT-PCR Master Mix
- Manufactured under ISO 13485:2016 for research use only
- CE-IVD Pending
Detect SARS-CoV-2 specific RNA with real time RT-PCR According to the Charite/Berlin Protocol
Figure 1. Duplex TaqMan RT-PCR analytical sensitivity for each of the SARS-CoV-2 targets based on the Charité/Berlin protocol. The analytical sensitivity was determined by analyzing six serial dilutions (200,000 copies to 2 copies) of Norgen’s positive control from the COVID-19 TaqMan RT-PCR Kit (E/RdRP gene)(Cat. TM67200). Five µL of template was used in a 20 µL of the one-step RT-PCR reaction and the arrow indicates a copy number of 10.
| Cat. # | Description | Size | |
|---|---|---|---|
| Cat. TM67200 | COVID-19 TaqMan RT-PCR Kit (E/RdRP genes) | 500 rxns | View product page |
Option 2: 2019-nCoV TaqMan RT-PCR Kit
| Cat. # | Description | Size | |
|---|---|---|---|
| TM67100, TM67120 | 2019-nCoV TaqMan RT-PCR Kit | 50 rxns, 500 rxns | View product page |
+ PCR COMPONENTS
2019-nCoV Primer & Probe Mixes
- COVID-19 detection based on the assays and protocols
developed by the CDC - Primer & Probe Mixes contain all 3 CDC developed assays in individual tubes
- All assays are premixed to the working concentrations
recommended by the CDC - Positive Control contains 2 nCoV nucleocapsid target gene RNA (N1,N2) and RNase P (internal control) which are compatible with the CDC primer/probe set
- Convenient ready-to-use 2X One-Step RT-PCR Master Mix
- Manufactured under ISO 13485:2016 for research use only
Ordering Information
| Cat. TM67101 | 50 Rxns |
| Cat. TM67130 | 500 Rxns |
2019-nCoV RT-PCR Positive Control
PositiveControl(Cat.PC67102)-TheoreticalCopyNumber
| 2019-nCoV_N1F | 2019-nCoV_N2F | RNAse P-F | |
|---|---|---|---|
| 2×108 | + | + | + |
| 2×107 | + | + | + |
| 2×106 | + | + | + |
| 2×105 | + | + | + |
| 2×104 | + | + | + |
| 2×103 | + | + | + |
| 2×102 | + | + | + |
| 2×101 | + | + | + |
| NTC | N/D | N/D | N/D |
- Positive control for SARS-CoV-2, which causes COVID-19 (2019-nCoV)
- Contains 2 nCoV nucleocapsid target gene RNA (N1, N2) and RNase P.
- Compatible with CDC primer/probe set
Ordering Information
| Cat.PC67102 | 50 µL |
| Cat.PC67120 | 500 µL |
2X One-Step RT-PCR Master Mix
- Convenient ready-to-use solution
- High sensitivity and yield
- Robust amplification
Ordering Information
| Cat. 28113 | 100 Rxns |
| Cat. 28114 | 200 Rxns |
| Cat. 28115 | 500 Rxns |
Figure 1. Performance of Norgens 2X One-Step RT-PCR Master Mix (28113) to detect SARS-CoV-2 using Norgens 2019-nCoV Primer & Probe Mixes (TM67101) and Norgens 2019-nCoV RT-PCR Positive Control (PC67102). The one-step RT-PCR reaction was prepared according to the CDC protocol and the sensitivity of detection of the three targets; 2019-nCoV_ N1 (Circle), 2019-nCoV_ N2 (Triangle) and RNase P (Cross), was compared with Competitor Ns product (Red).