Products using the MultiCode®-RTx Technology from Luminex offer a flexible platform for both real-time and multiplex PCR-based assays. MultiCode products are based upon the unique MultiCode bases, isoC and isoG. The synthetic isoC:isoG DNA base pair differs from the naturally occurring base pair in its hydrogen bonding pattern. As a result, the MultiCode bases, isoC and isoG, can only base pair with each other. This property enables site-specific incorporation of the isobases during amplification.
The isoC and isoG MultiCode bases form the building blocks for Luminex’s next generation MultiCode assays for nucleic acid-based testing. MultiCode-based PCR assays are used for the early detection of infectious diseases and genetic-based conditions.
The reporter-labeled forward primer containing a single isoC on the 5’ end and unlabeled reverse primer hybridize to the target nucleic acid. During the amplification process, the labeled primer is incorporated into the newly synthesized strand and serves as template for the reverse primer in the next cycle.
Synthesis of the opposite strand terminates with the incorporation of an isoG with a covalently attached quencher molecule. The resulting proximity of the quencher to the reporter produces a decrease in fluorescence. The fluorescence decrease is directly proportional to the amount of amplicon.
Following the completion of amplification, a thermal melt is performed and fluorescence is restored after the strands separate.
Fluorescence is monitored on commonly available real-time thermal cyclers. Process controls may be utilized to monitor extraction and amplification. The MultiCode-RTx Analysis Software1 allows the user to import raw data generated from different real-time instruments. The software also features the ability to automatically graph standard curves, perform quantitative calculations and generate custom reports.
MultiCode®-RTx Technology — Product Information
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