The combination of highly stringent padlock probe-dependent rolling circle amplification technique and sensitive lateral flow biosensors enable simultaneous detection of multiple targets. Several bacterial and/or viral species, both wild-types and mutants, as well as other single nucleotide variants can be conveniently detected in a single test.
NOVEL LFA-DNA INTEGRATION
Our novel diagnostic method consists of integrating existing lateral flow assay-technology with DNA oligonucleotides, which in conjunction with our amplification kit provides a sensitive method for detection of pathogenic genetic content.
The detection of the pathogenic genetic content is done by using gold-conjugated oligonucleotides that attach to either the pathogenic genetic content, or to a control tag, which is attached to the surface of the LF-sheet. The target sequence (pathogenic genetic content) is in turn bound to a test tag, which is attached to the surface of the LF-sheet. The gold-conjugated oligonucleotides will be visible to the naked eye a sufficient amount of target sequences have been bound.
State of the art DNA amplification. Novel DNA integration.
LINEAR, ISOTHERMAL DNA AMPLIFICATION
In order to achieve the high sensitivity we require for our products, an amplification step is performed. Our technology is based a patented version of the RCA (Rolling Circle Amplification) technology, which makes it possible to perform an isothermal amplification of a target sequence. This can be seen in (A), in the figure below.
First, a PLP (padlock probe) is created (grey, below), which is designed to have complementary 5' and 3'-ends against the target sequence (orange, below). If a match occurs, ligation can be performed and the amplification itself can start. The linear tandem repeat sequence is called an RCP (Rolling Circle Product), and it is monomerised in the following step. This allows for an even greater amplification by performing an additional subsequent RCA.
After the last monomerisation, the monomers are flown through the LF-membrane, and the detection itself takes place. This can be seen in (B), in the figure below.
Bordering on antibiotic resistance, EMPE’s product line also follows identification of various cancerous tissues, zoonotic and veterinary infection diagnostics, and antibiotic abuse in food and beverages.