Pathogen/Viral Nucleic Acid Isolation Kit (Magnetic Bead System)

Aperçu du produit

Norgen’s Pathogen/Viral Nucleic Acid Isolation Kit (Magnetic Bead System) is designed to extract nucleic acid (DNA and RNA) from viruses and easily lysable bacteria found in body fluids and transport mediums. The purification is based on Norgen’s magnetic bead-based technology that is designed for efficient high throughput extraction of high-quality DNA as well as RNA in a single elution. The kit is compatible with automation on NorMag purification system (Cat. 72715, Cat.72705), KingFisher™ Purification Systems or other similar systems, as well as liquid handling systems such as Hamilton Star/Vantage or Tecan.

The Pathogen/Viral Nucleic Acid Isolation Kit (Magnetic Bead System) allows extraction of DNA and RNA from up to 200 µL of different bodily fluids and transport mediums such as saliva, urine, blood, plasma, VTM, Norgen’s Total Nucleic Acid preservative and more. An optional step in the protocol also allows for the isolation of nucleic acids from gram positive bacteria by incorporating a lysozyme incubation. The nucleic acids are preferentially purified from the cellular proteinaceous components, followed by binding to the magnetic beads and co-elution of nucleic acids in an elution volume of 50-100 L. The purified DNA and RNA is of the highest integrity and can be used in a number of downstream applications including real time PCR and NGS applications.

Détails

Supporting Data

Figure 1. Multiple viral pathogens were detected in clinical samples with Norgens Ct values being significantly lower than those when a competitors kit was used. Nasopharyngeal swabs were taken at a hospital in Southern Ontario in January 2024. Swabs were immediately stored in a transport medium, sample_2 and sample_8 in McMaster Molecular Medium (MMM) and the other six samples were stored in Copan UTM. A multiplex qPCR system was used that tested for 10 targets and an internal control. The 10 targets include strains of Adenovirus, Influenza A/B, MPV, Covid, Parainfluenza, and RSV. The reactions were run on a Rotorgene Q machine.

Figure 2. qPCR for viral pathogens (COVID N1/N2) and bacterial pathogens (16S/S. aureus) shows that pathogen nucleic acids can be extracted and detected from a variety of bodily fluids. 200 μL of each sample (blood, plasma, saliva, urine) was spiked with COVID particles and Nucleic Acids were extracted using Norgen’s Pathogen/Viral Nucleic Acid Isolation Kit. After the extraction qPCR was performed using 2 μL of eluate in a 25 μL PCR reaction or 5 μL of eluate for one step RT-PCR reaction.

Figure 3. qPCR for viral pathogens (COVID N1/N2) and bacterial pathogens (16S/S. aureus) shows that pathogen nucleic acids can be extracted and detected from a variety of transport media. 200 μL of each transport medium (Norgen’s Total Nucleic Acid Preservative, PBS, Zymos Shield, VTM) was spiked with COVID particles and Nucleic Acids were extracted using Norgen’s Pathogen/Viral Nucleic Acid Isolation Kit. After the extraction qPCR was performed using 2 μL of eluate in a 25 μL PCR reaction or 5 μL of eluate for one step RT-PCR reaction.

Figure 4. qPCR of DNA (GAPDH) and RNA (RNaseP) housekeeping genes shows that both DNA and RNA can be extracted from a variety of bodily fluids. 200 μL of each sample (blood, plasma, saliva, urine) was spiked with COVID particles and Nucleic Acids were extracted using Norgen’s Pathogen/Viral Nucleic Acid Isolation Kit or the MagMAX™ Viral/Pathogen Nucleic Acid Isolation Kit. After the extraction qPCR was performed using 2 μL of eluate in a 25 μL PCR reaction or 5 μL of eluate for one step RT-PCR reaction.

Figure 5. qPCR of DNA (GAPDH) and RNA (RNaseP) housekeeping genes shows that both DNA and RNA can be extracted from a variety of transport media. 200 μL of each transport medium (Norgen’s Total Nucleic Acid Preservative, PBS, Zymos Shield, VTM) was spiked with COVID particles and Nucleic Acids were extracted using Norgen’s Pathogen/Viral Nucleic Acid Isolation Kit or the MagMAX™ Viral/Pathogen Nucleic Acid Isolation Kit. After the extraction qPCR was performed using 2 μL of eluate in a 25 μL PCR reaction or 5 μL of eluate for one step RT-PCR reaction.

Figure 6. qPCR curves showing amplification of the COVID N2 transcript from Norgen's Total Nucleic Acid Preservative. 200 μL of each transport medium (Norgen’s Total Nucleic Acid Preservative, PBS, Zymos Shield, VTM) was spiked with COVID particles and Nucleic Acids were extracted using Norgen’s Pathogen/Viral Nucleic Acid Isolation Kit (turquoise) or the MagMAX™ Viral/Pathogen Nucleic Acid Isolation Kit (royal blue). After the extraction qPCR was performed using 2 μL of eluate in a 25 μL PCR reaction or 5 μL of eluate for one step RT-PCR reaction.

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Pathogen/Viral Nucleic Acid Isolation Kit (Magnetic Bead System)