PlexZyme®

High performance and reliable multiplex qPCR detection. PlexZyme® is the driver behind the entire SpeeDx product portfolio.

Sensitive and Quantitative

Sensitive and Quantitative

Can detect less than 10 copies of target and has a good dynamic range from 106 to 10 copies.
Highly Specific

Highly Specific

Specificity is determined by 5 binding events of the 2 primers and 2 partzymes to the amplicons, plus probe for signal generation.
Flexible

Flexible

Probe design is independent of target sequence, allowing for the use of a suite of well-characterised universal probes.

PlexZyme® technology offers high performance and reliable qPCR detection and is the driver behind the entire SpeeDx product portfolio. During PlexZyme® qPCR, primers amplify target nucleic acid sequences and produce amplicons which serve as a template for PlexZyme® formation. Once the partzymes have assembled into PlexZyme® enzymes, universal probes bind and enzymatic cleavage of the probes between fluorophore and quencher dye pairs generates fluorescence. Changes in fluorescence allow detection and/or quantification of the target nucleic acid in real time.

PlexZyme binding to target and cleaving a probe

PlexPrime®

Novel nucleic acid amplification approach, creating amplicons distinctly different from the parent sequence. The combination of PlexPrime® and PlexZyme® technology enables multiplex mutation detection with high sensitivity and specificity.

Multiplex compatible

Multiplex compatible

Detect multiple mutations in a single reaction.
Versatile

Versatile

Flexibility for challenging sequences.
Highly Specific

Highly Specific

Increased stringency due to insert sequence

The PlexPrime® primer is composed of three regions:

  • 5’ region anchors the primer to a particular location in the target
  • 3’ region selectively amplifies the target sequence of interest
  • the Insert, acting as a bridging structure which both (a) inserts target-independent sequence into the resulting amplicon and (b) increases the selective pressure of the 3’ region

 

The unique feature of the PlexPrime® primer make it extremely suitable for detection and discrimination of mutations or Single Nucleotide Polymorphisms (SNPs) in a multiplex reaction or a pool of nucleic acid sequences containing non-conserved regions.

PlexZyme binding to target and probe

References

Mokany E., Tan Y.L., Bone S.M., Fuery C., Todd A.V., 2013. MNAzyme qPCR with superior multiplexing capacity. Clin. Chem. 59(2); 419-426

Link | Article | Supplementary Information

Mokany E., Bone S.M., Young P.E., Doan T.M., Todd A.V., 2010. MNAzymes are a versatile new class of nucleic acid enzymes which can function as biosensors and molecular switches. J. Am. Chem. Soc. 132(3); 1051-1059

Link Article | Supplementary Information

Tan L. Y., Walker S. M., Lonergan T., Lima N. E., Todd A. V. Mokany E. 2017. Superior Multiplexing Capacity of PlexPrimers Enables Sensitive and Specific Detection of SNPs and Cluster Mutations in qPCR. PLOS One January 23rd.
Link

Contact SpeeDx for more information about these diagnostic products.