PyroMark™ID

Sequence and identify 96 isolates in 1 hour

Biotage announces a complete solution for sequence-based microbial identification and resistance typing: PyroMark™ID.

Sequence data has always been the Gold Standard for microbial identification, but the time, effort and uncertainties associated with previous solutions have hindered routine application. PyroMark ID is based on Pyrosequencing^®, the only genetic method capable of generating real sequence data in minutes.

PyroMark ID is designed for routine microbial identification for clinical research. PyroMark ID supports microbial identification by streamlining the entire process from assay design and sequencing through to the discrimination of the microbial species. PyroMark ID integrates the sequencing process with the interpretation of Pyrosequencing data, using the new identification software IdentiFire™. IdentiFire compares the sequence data generated by Pyrosequencing against a local database of relevant microbial species.

PyroMark ID exploits the improved sequencing performance attained by Pyro Gold, the new chemistry formulation for Pyrosequencing. Pyro Gold improves sequence read length and quality as well as first-time success in assay design.

PyroMark ID also supports sequence-based resistance typing, by analysing SNPs, point mutations, insertions, deletions and multiple gene copies on the same instrument platform.

 Read more about Pyrosequencing in Microbiology

PyroMark ID, Microbial Identification by Pyrosequencing

See how PyroMark ID generates sequence data and IdentiFire matches the data to a local database.

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How PyroMark ID works, step-by-step (approximate time in brackets)

			Perform a PCR reaction to amplify microbial DNA (RT-PCR for viral RNA). One of the PCR primers is biotinylated in order to create a single-stranded DNA template in the next step (30 minutes - 2 hours).
		Create single-stranded DNA from the PCR product using the Vacuum Prep Workstation and Tool (15 minutes).
			Run the Pyrosequencing reaction on the single-stranded DNA template. Nucleotides are dispensed in a cyclic manner, and each incorporation event generates a flash of light proportional to the number of nucleotides incorporated, displayed as a peak in the Pyrogram™. The sequence is base-called automatically (1 minute/nucleotide; up to 96 samples in parallel).

IdentiFire™ software compares the generated DNA sequence to a local database. The database contains DNA sequences that act as signature sequences for specific microbes, which can be either obtained from public sources or from the user’s own validated sources (set up and analyze 96 samples in 1 minute).

			IdentiFire™ software makes a full report of identified species, with confidence scores. IdentiFire performs the analyses and reports are generated. In this example, Staphylococcus aureus was identified.