Fire Wolf by RevoluGen - DNA amplification technology

 

Milo is a 55 nucleotide recognition sequence

Milo is a very accurate DNA diagnostic technology. Milo is so accurate that it can detect a one base pair difference between test samples. This patented chemistry has multiple advantages over its nearest competitor the Molecular Beacon DNA identification system.

 Fundamentally, the Milo sequence recognition portion is nearly twice the length of the Molecular Beacon system as illustrated in the following diagram. Stability problems tend to limit the Molecular Beacon system to identification sequences of between 25 to 30 base pairs long. The Milo identification sequence is stable up to 55 base pairs long. This greatly improves the accuracy of the recognition system.

 A schematic of the Milo technology compared to the Molecular Beacon system

Revolugen-Milo-55-base-pair-diagram-Feb-18

Milo also has a number of other features that make it attractive as a component in a molecular diagnostic system, these include;

No genomic limitations

There are no thermodynamic restrictions that restrict Milo from being designed to work on any part of the genome. The Molecular Beacon system does not work on all of the genome because of its required operating paramaters. This is important as it very usefully enables Milo to be designed to work anywhere on the genome.

In-built error reduction

The underlying chemistry within Milo includes an automatic degree of self monitoring and therefore error reduction. This is part of the very high accuracy rate (single base pair differentiation) that Milo produces.


Multiplexing facility

The accuracy of the underlying chemistry also facilitates Milo’s ability to be multiplexed.


Specificity

Milo can be very accurate and detect differences down to a single base pair difference in its identification sequence. This identification signal has been further improved as the following test data illustrates. Under RevoluGen’s optimisation process the signal strength from Milo was more than tripled for a single base pair difference.

Assay test results using Firew Wolf by Revolugen
Specificity charts for MILO Revolugen NAIP kit
A chart showing Neisseria gonnorrhoea (NG) molecular testing using Fire Wolf by Revolugen
Revolugen Base pair recognition differentation.

Sensitivity

Milo is sensitive enough to be seen to identify as few as XXXX molecules with present reporter optics. In practical terms this translates into some 10 cells when allied to a suitable amplification system as illustrated below.

Fire Wolf RNA technology by revolugen

Interference resistance

Milo remains effective (when allied with a suitable amplification system) even in the presence of overwhelming contamination for other bacteria in the sample as the test results detailed above show.