BART
The ability to combine the power of molecular detection technologies with simple to use, robust and low-cost hardware is what differentiates Lumora's technology from competing systems. This represents a new paradigm in molecular diagnostics where expensive, complicated hardware & consumables have excluded many potential users.
In essence, Lumora's technology makes the sample do all the work rather than the hardware or user: that's how Lumora's BART technology makes molecular diagnostics easy.
BART (Bioluminescent Assay in Real-Time) is our novel reporter system that permits real-time, quantitative detection of nucleic acids during isothermal DNA amplification without the need for fluorescent reporters.
With BART, the sample does NOT need to be irradiated, visually inspected or form part of an electrochemical circuit.
Using BART is easy as the BART reagents are simply mixed with the amplification reagents used. No specialist consumables are required and the hardware used to perform BART-based assays is the simplest yet in molecular diagnostics.
Key features of using BART reagent as a reporter system include:
- Quantitative, real-time output
- Closed-tube format, preventing amplicon carry-over
- Can decrease the time-to-result compared to other reporter systems
- Proven with several isothermal amplification technologies
- Can tolerate the presence of magnetic beads (and other particulates) within the sample
- No specialist consumables required
As a consequence of the above, remarkably simple hardware can be used to perform BART-based assays. This simplicity means that very robust and low-cost devices can be manufactured for use in challenging settings which have traditionally rejected molecular methods as too costly and fragile for their needs. As such, BART is an enabling technology for the point-of-use molecular market in particular.
However, the simplicity of BART also means that with very simple hardware (just a heating block and a CCD camera) hundreds of samples can be processed simultaneously, in real-time. Hence BART-based systems also represent a means for increasing throughput in testing laboratories without the requirement for complicated and costly hardware systems.
The basis of BART technology
BART works by coupling the generation of pyrophosphate, a by-product of nucleic acid synthesis, to the emission of light from a highly thermostable version of firefly luciferase (figure 1). As such, amplification of a specific target nucleic acid sequence becomes linked to a luminescent output from the sample itself.
BART produces light outputs that give both a rapid increase and decrease in light only when a target sequence is amplified (figure 2).
This facet of the BART output has several advantages in diagnostics including the fact that very simple algorithms can be employed to quickly ascertain whether a sample is positive or not. This can be utilised in devices where the user can be informed as early as possible of whether an important sample is positive or not (e.g. figure 3).
Lumora has shown that incorporating the BART reagents into an amplification reaction has no adverse affects on amplification. In fact, it is possible to achieve faster kinetics with BART compared to certain fluormetric techniques. As such, rapid, specific, sensitive and quantitative results can be obtained with a suitable isothermal amplification method (figure 4, showing two dilution series of target nucleic acid with different Nucleic Acid Amplification Technologies, 'NAATs').
Lumora is presently developing two very different hardware systems for use with the BART technology. One of these based on the use of a CCD-Camera and is capable of very high throughput in laboratory settings; the other is a low throughput device (8-24 samples) designed for point-of-use applications in a variety of settings (figure 5).
Both the CCD-Camera and photodiode system can take advantage of the unique outputs from BART where it is the rate of change of light emission, rather than absolute light intensity, that is the key factor. This significantly simplifies the hardware requirements as inter-sample calibration for absolute light output is unnecessary.
Presently, the perception of molecular diagnostics is that of an expensive, laboratory-bound, complicated process. With BART this perception is set to change, a new generation of users will realise that molecular can be made easy.
To download further information on BART, click here