2SmartFinder®: Multiplex PCR Technology Based on Melting Curve Analysis

2SmartFinder

The 2SMARTFinder® technology is based on the SmartFinder® technology, which is PathoFinder’s proprietary state-of-the-art multiplex PCR technology for Real Time Molecular Diagnostics. It enables Single tube Multiplex Amplification in Real Time (SMART) of up to 22 infectious pathogens in a single clinical sample.

In the 2SmartFinder technology, a pre-amplification step using target specific primers is followed by a signal amplification step using a for all targets a single amplification primer set. The pathogen identification is based on Melting Curve Analysis in different detection channels (FAM, ROX and Cy5) and uses non-target related specific sequences which are incorporated in the generated amplicons.

 

Despite the developments in conventional PCR, the complexity of multiplex Real Time PCR is still limited due to the lack of sufficient detection channels. To achieve high-end multiplexing capacity on standard Real Time PCR machines, PathoFinder has developed the SmartFinder® technology. For every SmartFinder® reaction, up to 12 targets can be identified by means of melting curve analysis. After a gene-specific pre-amplification step a signal amplification step follows that makes use of a single PCR primer pair for all targets, of which one primer is labelled with a  fluorescent dye (FAM). The detection of the amplified FAM labelled probes is by melting curve analysis on Real Time PCR systems. Twelve detection  probes, either ROX or Cy5 labelled and varying in melting temperature,  enable specific detection of the amplified product and the corresponding pathogen. The Internal Control (IC), which is added at the start of the procedure, is detected by a specific detection probe in an additional channel to validate a negative sample result. Our next generation 2SmartFinder assays return results within 2.5 hours after nucleic acid extraction for both respiratory and gastro-enteritis related pathogens, including viruses, bacteria and parasites. The 2SmartFinder technology is universally applicable and is used in the following products:

RespiFinder® 2SMART

50 Assays Cat. No. PF2600-2S

MeningoFinder® 2SMART

50 Assays Cat. No. PF1200-2S

GastroFinder® 2SMART

50 Assays Cat. No. PF5600-2S

STD-Finder® 2SMART

50 Assays Cat. No. PF3200-2S