TwistDx

Technology

TwistDx’s technology, RPA (Recombinase Polymerase Amplification), represents a complete evolution in diagnostics, combining superiority in speed, , portability and accessibility with exquisite sensitivity and specificity versus other diagnostic testing systems now on the market. It will be usable in nearly any setting, eliminating the need of a trained technician or laboratories.

TwistDx’s RPA process employs enzymes, known as recombinases, which are capable of pairing oligonucleotide primers with homologous sequence in duplex DNA. Through this method, DNA synthesis is directed to defined points in a sample DNA. If the target sequence is indeed present, an amplification reaction is initiated; no other sample manipulation such as thermal or chemical melting is required. The reaction progresses rapidly and results in specific amplification from just a few target copies to detectable levels within 15 minutes. The entire reaction system is stable as a dried formulation and can be transported safely without refrigeration.

Specific benefits of the RPA technology include:

Portability/Point of Care
RPA operates at low, constant temperatures and does not require initial melting of the sample DNA (optimal temperature 37°C); in fact, body heat can support the process if necessary. It is also fully robust to off-temperature and to low temperature set-up. At typical ambient temperatures (25°C) the process works, albeit more slowly, and results can still be obtained in 45-90 minutes – we anticipate completely disposable tests in the near future.
Speed
RPA is capable of amplifying from a few molecules to detectable levels within 15 minutes (at around 37°C). Combining this performance with the very simple rapid sample preparation and decentralized point-of-use device capability offers the potential for diagnosis within a half hour compared to the 24-hour turnarounds currently typical today for clinical samples due to a need to send them to centralized labs.
Sensitivity
RPA can detect single copies of DNA and tens of copies of RNA (or less).
Specificity
RPA is so specific that it can operate easily to single molecule levels in the presence of hundreds of nanograms of unrelated complex genomic DNA such as that from plants and mammals. This permits the detection of trace levels of targets even in extremely complex nucleic acid samples.
Broad Applicability
RPA can be readily applied to any DNA or RNA target. Ultra-high sensitivity RNA detection is possible in a one-pot system if reverse transcriptase in included.
Simultaneous Multiple Target Detection
Several different potential targets may be detected with a single RPA test as multiple detection primers can be combined in one tube.
Ease of Use
In multiple validated cases, RPA has been shown to not require a clean sample (such as those generated by single step lysis procedures), and thus will ultimately be available for portable systems used by untrained personnel.
Low Upfront Cost Burden
Minimal or no device requirements broaden users compared to the restricted highly capitalized environments today.
No Refrigeration
The reaction system can be stabilized in a dried format permitting transportation and storage without refrigeration.

Sensitivity
RPA can detect single copies of DNA and tens of copies of RNA (or less).

Specificity
RPA is so specific that it can operate easily to single molecule levels in the presence of hundreds of nanograms of unrelated complex genomic DNA such as that from plants and mammals. This permits the detection of trace levels of targets even in extremely complex nucleic acid samples.

Broad Applicability
RPA can be readily applied to any DNA or RNA target. Ultra-high sensitivity RNA detection is possible in a one-pot system if reverse transcriptase in included.

Simultaneous Multiple Target Detection
Several different potential targets may be detected with a single RPA test as multiple detection primers can be combined in one tube.

Ease of Use
In multiple validated cases, RPA has been shown to not require a clean sample (such as those generated by single step lysis procedures), and thus will ultimately be available for portable systems used by untrained personnel.

Low Upfront Cost Burden
Minimal or no device requirements broaden users compared to the restricted highly capitalized environments today.

No Refrigeration
The reaction system can be stabilized in a dried format permitting transportation and storage without refrigeration.

Sensitivity RPA can detect single copies of DNA and tens of copies of RNA (or less).

Specificity RPA is so specific that it can operate easily to single molecule levels in the presence of hundreds of nanograms of unrelated complex genomic DNA such as that from plants and mammals. This permits the detection of trace levels of targets even in extremely complex nucleic acid samples.

Broad Applicability RPA can be readily applied to any DNA or RNA target. Ultra-high sensitivity RNA detection is possible in a one-pot system if reverse transcriptase in included.

Simultaneous Multiple Target Detection Several different potential targets may be detected with a single RPA test as multiple detection primers can be combined in one tube.

Ease of Use In multiple validated cases, RPA has been shown to not require a clean sample (such as those generated by single step lysis procedures), and thus will ultimately be available for portable systems used by untrained personnel.

Low Upfront Cost Burden Minimal or no device requirements broaden users compared to the restricted highly capitalized environments today.