COVID-19 TaqMan RT-PCR Detection Kit (N/ORF1ab genes) Dx

Cat. DxTM67300

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COVID-19 TaqMan RT-PCR Detection Kit (N/ORF1ab genes) Dx

Cat. DxTM67300

CE-IVD marked in accordance with EU Directive 98/79/EC
Multiplex RT-PCR system detects two SARS-CoV-2 specific genes (N gene and ORF1ab) in a single one step RT-PCR reaction
Primer & Probe Mixes contain all N gene, ORF1ab and human RP assays
Positive Control contains the three SARS-related target genes: N gene, ORF1ab gene as well as the human RP (internal control).
Convenient ready-to-use 2X One-Step RT-PCR Master Mix
Find out more information about our complete COVID-19 workflow

Intended for in vitro diagnostics use.

CE marked version is not available for sale in the US and Canada

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COVID-19 TaqMan RT-PCR Kit (N/ORF1ab genes) Dx
(Cat. DxTM67300) 500 rxns

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Product Format
Norgen Biotek offers kits in different standardized formats. You can view and compare kit components in the Components Table in the Product Overview section. For more information on format types, please visit our Format Information page.

Product Size

Kit Specifications

You have selected: Cat. DxTM67300

Storage Conditions and Product Stability

All kit components should be stored at -20°C upon arrival
Repeated thawing and freezing (> 3x) of the Master Mix and Positive Control should be avoided, as this may affect the performance of the assay. If the reagents are to be used only intermittently, they should be frozen in aliquots.
All reagents can be stored for 6 months at -20°C without showing any reduction in performance.

Supporting Data

Figure 1: Multiplexed RT-PCR using the COVID-19 TaqMan RT-PCR Kit (N/ORF1ab genes) Kit for the specific detection of SARS-CoV-2 (N gene and ORF1ab) and an internal human control gene (RNase P). Shown are exemplary amplifcation curves following RT-PCR of 5 µL of the provided Positive Control (n = 3).

Figure 2: Highly sensitive and reproducible SARS-CoV-2 viral particle detection from varied sample matrices. Serial dilution of quantified heat-inactivated SARS-CoV-2 viral particles were spiked into preserved Nasal, Oral, and Saliva samples prior to RNA isolation using Norgens Saliva/Swab RNA Purification Kit (Cat. 69100) and RT-PCR to detect SARS-CoV-2 N gene and ORF1ab, and human RNase P (internal control). A) Linear detection of down to ten (10) viral copies per PCR reaction was reliably achieved in all sample types (n = 3; error is standard deviation).

Figure 3: Highly specific detection of SARS-CoV-2; No cross reactivity with other similar/major pathogens. Pathogen concentrations of ≥ 105 pfu/mL (for viruses), ≥ 106 CFU/mL (for bacteria), or 200,000 copies/uL of COVID-19 TaqMan RT-PCR Kit (N/ORF1ab genes) Positive Control were spiked into Nasal Swab and Saliva samples prior to RNA isolation using Norgens Saliva/Swab RNA Purification Kit (Cat. 69100) and RT-PCR to detect SARS-CoV-2 N gene and ORF1ab, and human RNase P (internal control). Positive SARS-CoV-2 detection was achieved only from SARS-CoV-2-spiked and Postive Control-spiked samples.

Figure 4: Robust detection, even with common sample contaminants. Nasal Swab and Saliva samples were supplemented with common contaminents either following or during collection, with a select subset shown here. 4 x LoD (40 viral copies per PCR reaction) of heat-inactivated SARS-CoV-2 viral particles were spiked into samples prior to RNA isolation using Norgens Saliva/Swab RNA Purification Kit (Cat. 69100) and RT-PCR to detect SARS-CoV-2 N gene and ORF1ab, and human RNase P (internal control). Positive SARS-CoV-2 detection was achieved in all cases (n = 5; error is standard deviation).

Figure 5: RT-PCR data interpretation key. In addition to this, the No Template Control (NTC) must not exhibit fluorescence above threshold; and each target in the N/ORF1ab/RP Positive Control must be detected (ideally <25 Ct). If these conditions are not met, the data is invalid, and the assay must be repeated.

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Protocols:

(DxTM67300) COVID-19 TaqMan RT-PCR Kit (N/ORF1ab genes) Dx - Protocol

Product Overview
Documentation

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COVID-19 TaqMan RT-PCR Detection Kit (N/ORF1ab genes) Dx

Norgen’s COVID-19 TaqMan RT-PCR Kit (N/ORF1ab genes) Dx includes 2X One-Step RT-PCR Master Mix, a primer/probe mix, a positive control and a negative control (nuclease-free water). The primer/probe mix contains three SARS-CoV-2 detection targets; N (Nucleoprotein), ORF1ab (Open reading frame1ab) and the human RNase P transcript (RP - HEX/VIC) as an internal control target to monitor for PCR inhibition, and to validate the quality of the sample and the detection result. The provided N/ORF1ab/RP Positive Control contains an in vitro RNA transcript for the two SARS-related target genes: N gene, ORF1ab gene as well as the human RP gene (internal control).

Positive results are indicative of SARS-CoV-2 RNA detection, however clinical correlation with patient history and other diagnostic information is necessary to determine patient infection status. Positive results do not rule out co-infection with other viruses and therefore the agent detected may not be the definite cause of disease. Negative results do not preclude SARS-CoV-2 infection and should not be used as the sole basis for patient management decisions. Any negative results must be combined with clinical observations, patient history, and epidemiological information.

Norgen’s COVID-19 TaqMan RT-PCR Kit (N/ORF1ab genes) Dx is intended for use by professional users including clinical laboratory personnel experienced and trained in molecular biology techniques including real-time PCR and in vitro diagnostic procedures.

Norgen’s COVID-19 TaqMan RT-PCR Kit (N/ORF1ab genes) Dx was developed and validated to be used with the BioRad CFX96 Touch^TM Real-Time PCR Detection System.