Chimerism monitoring

qPCR-based assay:

• KMRtype = Genotyping kit to select informative markers for chimerism monitoring
• KMRtrack = Monitoring kit to track chimeric status over time
• HLA-KMR = HLA loss detection assays

NGS-based assay:

• NGStrack = Chimerism monitoring by NGS (typing and monitoring)
• NGStrack-HLA loss = HLA loss detection assay (coming soon)

Software for analysis of chimerism data:

• KMRengine for analysis of KMRtype and KMRtrack data
• TRKengine for analysis of NGStrack (and coming soon: NGStrack-HLA loss data)

They were designed based on population data for European Caucasian, African, American Indian and Japanese populations. While we don’t have clear data for Asian subpopulations, these same set of populations proved well in the Korean population as published in The Journal of Molecular Diagnostics, Vol. 16, No. 6, November 2014 by Seon Young Kim et al. from the Department of Laboratory Medicine, Seoul National University College of Medicine, titled; Chimerism Monitoring after Allogeneic Hematopoietic Stem Cell Transplantation Using Quantitative Real-Time PCR of Biallelic Insertion/Deletion Polymorphisms.

The NGStrack assays have been designed based on known biallelic insertions and deletions. This

type of information is mostly known for the Caucasian population, which may therefore act as

the reference population. However, during our validation of the assays, different populations

have been tested and no bias towards a specific population has been detected.

KMRtype and KMRtrack are qPCR-based assays offering a sensitivity of up to 0.05% and a turnaround time of 2 hours (CE-marked). In contrast, NGStrack is an NGS-based assay with a sensitivity of up to 0.1% and turnaround time of 18-24 hours depending on sequencing time (RUO-marked).

All three assays use InDel markers, with significant overlap: 39 markers are shared by KMRtype and KMRtrack, while NGStrack includes 34 markers. The markers are shared between both methods allowing the user to easily switch between methods. With KMR the user must first run a genotyping experiment and select the informative markers to continue with KMRtrack, with NGStrack both genotyping and monitoring can take place in the same experiment as all markers are taken along the experiment set-up. With KMRtrack, a pre-sample is required for each monitoring sample, whereas NGStrack requires a pre-sample only during the initial typing experiment. With both assays more than 2 genomes can be analyzed allowing for the monitoring of chimerism after multiple transfers.

Each product has unique features and similarities, allowing you to select the best option tailored to your lab’s specific needs. Request more information via our Sales team.

The following instruments are currently compatible with KMRtype and KMRtrack Assays.

• ABI ViiA7 v1.2.4
• ABI 7500 Fast v2.0.6
• ABI 7500 v1.3.1/1.4/2.0.2/2.0.6/2.3
• ABI 7900HT Fast, v1.4 (AlleleSEQR chimerism only)
• Agilent AriaMx 1.5, 1.7
• AriaMx (1.5 & 1.7)
• Azure Cielo 6 (Azure Cielo manager)
• CFX Maestro software 4.1.2533
• qTower 3 (qPCRsoft 4.1)
• QuantStudio 5, v1.2, v1.4.3 and 1.5
• QuantStudio 6, v1.2/1.3
• QuantStudio 7, v1.2, v1.3
• QuantStudio 12K, v1.2
• QuantStudio DX, v1.2
• BioRad CFX 96, v1.6/3.0/3.1
• LightCycler 480 I, v1.5.0 (AlleleSEQR chimerism only)

The NGStrack libraries are intended for sequencing on an Illumina NGS platform. The NGStrack workflow is validated on the Illumina MiSeq platform and MiniSeq. Other Illumina sequencing platforms (iSeq, NextSeq) use similar chemistry, therefore the functional libraries generated with NGStrack workflow are anticipated to be compatible with all Illumina NGS platforms. Use of the NGStrack workflow on other Illumina platforms has to be validated by the user.

The following products are CE-IVD marked:

• KMRtype
• KMRtrack
• KMRassay
• KMRengine
• NGStrack and TRKengine will become available as CE IVD at a later timepoint

• The qPCR files generated using KMRtype/track or HLA-KMR can be analyzed with the software package KMRengine. This is a dedicated software package for qPCR chimerism analysis.
• The FastQ files generated using NGStrack can be analyzed with the software package TRKengine. This is a dedicated software package for NGS chimerism analysis.

Yes, TRKengine contains a CSV export that allows the transfer of results to LIMS.

• In KMRengine it is possible to export results in a PDF format.
• In TRKengine it is possible to export results in a PDF and CSV format.

• Via our website: gendx.com/support/send-us-your-files
• In KMRengine data can be sent through the menu option ‘Help’ > ‘Ask for Support’. This way log files and anonymized databases can be sent to our Technical Support team along with a question.
• In TRKengine data can be sent through the menu option ‘Support’ > ‘Ask for Support’. This way log files, anonymized databases and NGS data can be sent to our Technical Support team along with a question.

If not connected to the internet, copy the installer to the computer and use the ‘Activate offline’ option when following the on-screen instructions during installation. However, it is highly recommended to use ‘Online Activation’ to receive update notifications within the software.

Our Technical Support team is available if additional help is required.

This information can be found in the Instructions for Use. Or by contacting Sales.

The installers can be downloaded from this website. A license can be purchased via our Sales team.

The Certificates of Analysis of products or individual tubes can be downloaded from here.

Yes, the SDS can be downloaded from here.

• Contact Sales
• On our website: gendx.com/product_category/ngs-chimerism-monitoring
• Customers can find Instructions for Use here
• https://www.gendx.com/support/