RevoluGen’s second product is called Fire Wolf

Fire Wolf is a chemistry process that precisely identifies a target bacteria by looking at the 16S sequence of its ribosomal RNA. This 16S rRNA sequencing is the industry standard method for identifying all bacteria. Fire Wolf can detect differences as small as a single base pair change between similar bacterial 16S sequences. Fire Wolf is actually a proprietary combined, sequence amplification and precise identification technology – in other words it is our very accurate diagnostic test that also happens to be quite fast (about 30 minutes). With Fire Wolf we can design a specific diagnostic test for any bacteria in any Microbiome starting from just the knowledge of its 16S rRNA sequence alone; i.e. using its 16S rRNA base pair sequence as a digital barcode. Fire Wolf can also easily be multiplexed to allow for the identification of more than one bacterial type in a single test tube. Ease of design, lack of restricted genome areas and multiplexing are all considerable commercial advantages.

RevoluGen's Fire Wolf base pair recognition technology

This is a graphical representation of the steps in a generic Fire Wolf diagnostic Product. Panels of tests will be developed and multiplexed for individual target bacteria and clusters of different bacteria groups.    

Competitive Advantages

Revolugen NAIP technology 16S RNA / DNA

The company expects the Fire Wolf technology to generate multiple panels of in-vitro diagnostic tests for identifying many different bacteria in many different diseases. RevoluGen already has proof of concept for this idea’s first application. An initial version of this world-first test panel is expected to be available for out-licensing around mid 2018. A second stage version is expected to be the first of its kind to identify one meningitis causing bacteria without being confused by all the other innocent members of this family of bacteria. This diagnostic panel will be developed and ready for out-licensing around end 2018.

A Fire Wolf test is fast as it works inside a half hour, is accurate, as it can spot a single base pair difference between bacteria and it is cost effective, as each test can be multiplexed to do many tests at the same time in the same test tube. These features all combine to provide opportunities to generate panels of tests for whole groups of bacteria in the $2.2bn molecular diagnostics market. Fire Wolf may also eventually challenge many parts of the $17bn global infectious disease testing market.

Neisseria panel is the initial Fire Wolf target

The initial target for Fire Wolf is to develop a test panel for the Neisseria genus of bacteria. This test will identify the presence of every member of the complete family of bacteria that afflict humans. There are 13 bacterial members of this bacterial family that afflict humans, of which two species cause serious disease, but where the remaining eleven are innocent. The present tests on the market for the two bad bacteria are recognised to give false negative and false positive readings because of these confusingly innocent but similar commensal bacteria. Fire Wolf is so fast and accurate that it will be able to spot the bad from the innocent more precisely than all present technologies using a single multiplexed panel test. The first version of this panel is due to be available by for out-licensing around mid 2018.

Neisseria Gonorrhoea RNA DNA Revolugen NAIP

Following that, this principle of developing a multiplexed test panel covering a large number of the potential infectious diseases that can affect an organ could be developed further. For example, a specific Fire Wolf panel could then be developed to rapidly diagnose each of the well recognised infections that occur in the Urinary Tract, the Respiratory Tract, the Brain and in the Blood.

16S base pair sequence gel results by Revolugen
  • BASHH recommends the use of 2 molecular target sequences for identifying NG in extra-genital sites
  • We have identified 3 molecular target sequences on what is considered to be the gold standard for bacterial identification, the 16S rRNA sequence
  • Using the Leon technology, we can amplify three variable regions of the 16S rRNA sequences for discriminating NG from all other Neisseria species. These are the V1, V3 and V6 regions
  • Using the Milo technology, we can design a specific probe for members of the Neisseria genus in each of these three regions. Milo can be used to discriminate a single base mutation generating a differential signal of ~75% in comparison to the signal generated for the wild type region
  • This works on sequence areas where molecular beacons cannot be designed to work at all for any level of mutational analysis
  • In the Milo system, a single base difference equates to appx 3% sequence difference. 9% sequence difference generates a differential signal of ~97%. The V1, V3, and V6 regions show sequence differences of 3% right up to 45% between the NG, NM and all the other Neisseria species. This makes every species separately identifiable in any mix of species.
  • Stage 1 The first IVD panel incorporating this technology that RevoluGen will consist of a set of three independent diagnostic tests for NG. These tests will identify NG in any mix of all the other Neisseria species. There is currently no approved diagnostic test that has been approved for this application, with this detection power and for use in all body sites
  • Stage 2 The second IVD panel will identify NM in any mix of all the other Neisseria species. The Stage 2 panel will be based on a set of probes within 2 variable regions of the 16S rRNA (V1, V3)
  • NM in genital sites has been reported to be an emerging STI epidemic

Microbiome is the second Fire Wolf target

RevoluGen also thinks that Fire Wolf could become a key diagnostic tool for the bacteria that are important in the microbiomes that we live with. Many of these bacteria cannot be grown in the laboratory and are therefore not so well known. However, a Fire Wolf test panel can easily be designed for each bacteria once its 16S sequence is known. Fire Wolf is therefore one of the few technologies that could be used to commercialise rapid testing for the emerging microbiome markets. These microbiome markets could prove to become much bigger as the Microbiome & Genomic Testing is acknowledged as one of the “biggest game-changers in healthcare”.

It is well established that everything has its own microbiome, i.e. the mix of bacteria that live together as a community in and on every place of every body and product. These bacteria are responsible for much of the health and problems that afflict all living organisms. New tools such as the Next Generation Sequencing technologies are now beginning to identify all the different bacteria in each microbiome community. However, microbiomes are very complex and only just beginning to be understood. Some individual bacteria found in each microbiome are now beginning to be linked to specific health and disease issues. The ability to measure these key bacteria presents a multitude of commercial opportunities for Fire Wolf diagnostics.  


The present sequencing techniques that identify all these bacteria are accurate but too slow and expensive for consumer use. RevoluGen’s Fire Wolf technology is simple enough, fast enough and low cost enough, to permit its use as a consumer-friendly companion diagnostic for monitoring the specific good and bad bacteria within these microbiomes that clinical trials have shown to matter. With the relevant clinical knowledge about the important bacteria, foods and medicines can be designed try to manipulate these bacteria as steered by relevant diagnostic tests, in order to make people healthier. Fire Wolf diagnostics for these bacteria can be designed from just knowing the 16S rRNA digital sequences of these sort of bacteria, that often prove very difficult to grow in the laboratory.    

Management think that a low cost, companion diagnostics, such as a specifically tailored Fire Wolf panel of tests could also become critical to the commercialisation of any planned manipulation of the essential microbiomes associated with Functional Foods, Skin Care Cosmetics, Agriculture and Medicines for both humans and animals. Fire Wolf’s combination of speed, living bacteria-focus, accuracy of identification, test design-ease and multiplexing ability, opens-up a whole new range of breakthrough markets. RevoluGen is of the opinion that the growing understanding of the microbiome will lead to the need for consumer based diagnostic tests for the relevant bacteria.