Sensitive infectious disease testing

Intended Purpose

SwiftX™ ParaBact is designed for magnetic capture and sensitive and rapid extraction of bacteria as well as eukaryotic cells, i.e. parasitic protozoa, helminth eggs, fungi as well as human & animal cells.

Advantages

Improved cell capture

The cell capture buffer of SwiftX™ ParaBact has been optimized to enable sensitive binding of bacterial cells to our proprietary paramagnetic particles. As a consequence, the kit enables more efficient capturing of bacterial and parasites from liquid specimens compared to SwiftX™ DNA. This is truely convenient cell concentration - no centrifugation required, just use a magnetic rack. More details about which species we have tested, can be found in the product information material below.

Powerful cell lysis

With SwiftX™ ParaBact, the heat-assisted cell lysis that is typical for all SwiftX™ extraction kits, occurs under alkaline conditions. This leads to an improved rupture of bacterial cells, particularly of some gram-positive bacteria. The neutralization buffer ensures that the extract can be directly applied to downstream applications such as PCR, or isothermal amplifications. As a side effect of the alkaline conditions, RNA is actively degraded during cell lysis and will not influence your DNA assays.

Proven reverse purification

SwiftX™ ParaBact employs the same proprietary Reverse Purification technology as SwiftX™ DNA, which is based on our coated paramagnetic particles. Under lysis conditions, the coating of these particles allows for binding and active removal of inhibitors and cell debris from the cell lysate leading to a cleared lysate containing DNA of the lysed cells. Choose from our wide range of magnetic racks to ease your work.

For more information check out the FAQ section of SwiftX ParaBact.

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Applicable extraction targets

• Gram-positive and gram-negative bacteria
• Parasitic protozoa
• helminth tissue incl. eggs
• Fungi
• Human and animal cells

Applicable sample types

• Solid samples: swabs (buccal, nasal, pharyngeal, rectal, skin-lesions), pre-concentrated cells
• Liquid samples: blood, sputum, urine, cell cultures, saliva, fine-needle aspirates, tissue homogenates, cerebrospinal fluid, transport media, liquid-based cytology media, and wastewater.

Protocol 1

Direct extraction workflow incl. reverse purification:

• mix sample with Buffer BPL and Beads A
• incubate for 5 to 15 minutes at 95°C
• add Buffer AD to the lysate
• reverse purification by magnetic separation of Beads A
• use supernatant for downstream application

To be used with the following sample types:

• a pellet of pre-concentrated cells
• 50μL eluted swabbed sample (e.g. buccal, nasal, pharyngeal, rectal)
• up to 50μL of liquid

Protocols 2 and 3

Workflow for cell capture, extraction and reverse purification:

• add Buffer BPE and Beads A to liquid sample
• incubate for 3 minutes at room temperature
• place tubes in a magnetic rack
• remove and discard supernatant
• Protocol 3 only: wash Beads A once with Buffer BPE
• resuspend Beads A in Buffer BPL
• incubate for 5 to 15 minutes at 95°C
• add Buffer AD to the lysate
• remove tubes from heat block and place in a magnetic rack
• use supernatant for downstream application

To be used with the following sample types:

• Blood, urine, transport media
• Sputum, Saliva, CSF
• Homogenized tissue suspensions
• Liquid-based cytology medium
• Mammalian and bacterial cells in culture medium
• Wastewater

How is cell lysis different with SwiftX™ ParaBact?

The cell lysis procedure in SwiftX™ ParaBact and SwiftX™ DNA is essentially the same. It relies on heating the sample at 95°C. However, while in SwiftX™ DNA cell lysis is assisted by detergents only, in SwiftX™ ParaBact the lysis takes place under alkaline conditions, which improves the rupture of gram-positive bacteria without threatening the integrity of the DNA released during the lysis procedure. In addition, Buffer BPL also actively removes RNA from the lysate.

After lysis, neutralization of the samples needs to be carried out by adding Buffer AD. This renders the extracted DNA fully compatible with downstream applications such as PCR, LAMP or RAA detection.

What sample types can be extracted with SwiftX™ ParaBact?

SwiftX™ ParaBact offers three extraction protocols. The recommended buffer depends on the sample type to be analyzed.

For solid specimens such as swabs or pre-concentrated cells, we recommend Protocol 1.

If liquid samples shall be processed, e.g. blood, sputum, saliva, urine, tissue suspensions or cultured cells, then Protocols 2 or 3 are recommended as both include a cell capturing step preceeding the DNA extraction. The concentration of cells to a smaller volume contributes to the high sensitivity of DNA extraction with SwiftX™ ParaBact..

Protocol 3 is preferred if you intend to extract DNA from complex liquids such as whole blood or liquid-based cytology (LBC) media. In addition to the cell capture step, it includes a washing step before cell lysis in order to reduce the amount of inhibitory substances in the final DNA extract.

What is the function of Beads A in SwiftX™ ParaBact?

In SwiftX™ ParaBact, Beads A have the same function as in SwiftX™ DNA. You will find a detailed explanation in the FAQ section of SwiftX DNA.

Which cells or structures can be captured and enriched with Beads A from SwiftX™ ParaBact?

Buffer BPE and Beads A enable a centrifugation-free enrichment (cell capture) of various bacterial cells and tissue types. Similar to SwiftX™ DNA, SwiftX™ ParaBact was designed for magnetic capture of bacterial and eukaryotic cells, including parasitic protozoa, helminth eggs as well as human and animal cells. However, SwiftX™ ParaBact enables a species-independent capturing of bacterial cells over a wide range of different classes. A list of species tested for cell capturing can be found in the Technical Information in the download section above.

Do Beads A of SwiftX™ ParaBact bind nucleic acids?

No, Beads A are not capable of binding nucleic acids, neither RNA nor DNA. Beads A bind other biomolecules such as proteins and carbohydrates. Consenquently, they are used for clearance of the lysate after the heat extraction step by removing impurities such as cell debris and other inhibitory substances.