Virus Detection qPCR
Scientist using cryo tube for NAP and qPCR

Virus Detection: Nucleic Acid Purification and RT-qPCR

In SARS-CoV-2-research and vaccine development, a safe detection of the virus is mandatory. The current gold standard involves purifying the viral genomic material, followed by a reverse transcription-quantitative PCR (RT-qPCR), which has been proposed by the center of disease control1, and the World Health Organization2. In the first part of this process, the collected samples are usually aliquoted and reformatted to allow simpler handling. Processed viral genomes are then purified from the sample by nucleic acid purification (NAP). The released viral genomes are then used as an input for an RT-qPCR to detect different viral markers in the sample.

The methods and equipment shown are intended for molecular research applications. They are not intended, verified, or validated for use in the diagnosis of a disease or other human health conditions.

Collect Samples

Primary samples can be swabs, blood or other body fluids.

Storage and Management  

  • Samples can be stored in high quality Cryo tubes
  • An Ultra-Low Temperature Freezer allows long term storage of primary samples
  • The eLABJournal® and eLABInventory allow for effortless archiving and documentation 

Eppendorf CryoCube F570 ULT freezer for storage of lab samples More about Sample storage solutions

Aliquot Samples

To process the collected samples, they need to be transferred to tubes or plates.

Manual Liquid Handling

  • Mechanical or Electronic Single Channel Pipettes are ideal for precise reformatting of samples from primary vessels
  • Filter Tips assure the sample integrity of primary samples

More about Manual Liquid Handling


Automated Liquid Handling

  •  Benchtop Pipetting Robots support you to deal with an increased sample throughput

More about Automation Solutions

 


Plates and Tubes

  • Leachable free plates and tubes for optimal processing and storage of aliquoted samples

More about Laboratory Consumables


Storage and Management

  • Samples can be stored using high-quality Cryo tubes
  • Ultra-Low Temp Freezer allows long term storage of aliquoted samples
  • eLabJournal® and eLABInventory allow painless archiving and documentation

Eppendorf CryoCube F570 ULT freezer for storage of lab samples More about Sample storage solutions

Nucleic Acid Extraction

Nucleic Acid Extraction (also known as Nucleic Acid Purification) refers to the isolation of viral DNA or RNA used with magnetic-bead or filter/silica-based protocols.

Manual Liquid Handling

  • Single Channel Pipettes are ideal for processing in tube format. Multichannel pipettes are optimal for processing in plate format.
  • Filter Tips assure the sample integrity when handling primary samples
  • A Dispenser and Combitips® help add reagents in a quick and precise way. They also help minimize handling steps, used materials and reduce the risk of cross-contamination.

More about Manual Liquid Handling


Automated Liquid Handling

  • Benchtop Pipetting Robots allow walk-away automation of Magnetic-Bead-Based and Filter-Based Extraction Kits

More about Automation Solutions


Plates and Tubes

  • Leachable free plates and tubes for optimal results
  • A wide range of consumables allow you to pick the best-suited consumable for your protocol, ranging from single tubes to deep well plates
  • Safe and fast processing: easy one-hand handling, perfect fit on magnets, centrifugation and heat safe, automation-friendly

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Tempering and Mixing

  • ThermoMixers are especially helpful for lysis steps, but also allow to help with other incubation steps, like resuspension and neutralization
  • Suited for magnetic bead homogenization
  • Easily fits the biosafety cabinet
  • Interchangeable blocks make it easy to process and clean plates and tubes

More about Temperature Control and Mixing


Centrifugation

  • Key for processing filter-based protocols, especially single spin columns
  • A wide variety of centrifuges and adapters allow you to handle plates and single vessels of each size and format

More about Centrifugation


Storage and Management

  • Extracted nucleic acids can be stored in an Ultra-Low Temperature Freezer
  • eLabJournal® and  eLABInventory allow for effortless archiving and documentation

Eppendorf CryoCube F570 ULT freezer for storage of lab samples More about Sample storage solutions 

(RT-)qPCR setup

This step involves the preparation of reaction mixes and combining them with the samples for the qPCR analysis. If you use viral RNA, a reverse transcription (RT) step is involved here. This can be part of the qPCR process or a separate step depending on the used reagents and protocols. The quality of the qPCR setup has a big impact on the final results, which is why extra caution and concentration is required to perform this step.

Manual Liquid Handling

  • Single Channel Pipettes are ideal for processing samples in tube format. Multichannel Pipettes are optimal for processing in plate format
  • Superior precision for highest reproducibility and handling low volumes
  • A perfectly fitted tip for your pipette helps you to maximize precision
  • Filter Tips drastically reduce the risk of cross-contamination
  • A Dispenser and Combitips® help add reagents in a quick and precise way. They also help minimize handling steps, used materials and reduce the risk of cross-contamination.

More about Manual Liquid Handling

 


Automated Liquid Handling

  • Benchtop pipetting robots allow an effortless qPCR setup

More about Automation Solutions


Plates and Tubes

  • Leachable free plates and tubes avoid interference with the qPCR reaction
  • White plates help achieve maximum signal strength
  • The ultra-clear film is perfect to seal plates

More about Consumables


Centrifugation

  • Crucial to avoid air bubbles that interfere with the fluorescent signal and for spinning down samples and reagents
  • A wide variety of centrifuges and adapters allow you to handle plates and single vessels of each size and format

More about Centrifugation

(RT-)qPCR

The qPCR run helps to detect and quantify viruses, viral subspecies, or viral genes. Samples can be stored after the PCR for additional experiments.

Storage and Management

  • Plates and tubes can be stored using Ultra-Low Temperature Freezer
  • The eLabJournal® and eLABInventory allow for effortless archiving and documentation

Eppendorf CryoCube F570 ULT freezer for storage of lab samplesMore about Sample storage solutions

1 Interim guidelines for collecting, handling, and testing clinical specimens from persons under investigation (PUIs) for 2019 novel coronavirus (2019-nCoV).
Centers for Disease Control and Prevention, 2020
www.cdc.gov/coronavirus/2019-ncov/lab/guidelines-clinical-specimens.html
Accessed: 25.05.2020

2 Molecular assays to diagnose COVID-19: Summary table of available protocols
World Health Organization, 2020
www.who.int/docs/default-source/coronaviruse/whoinhouseassays.pdf
Accessed: 25.05.2020