Olerup News 151009 by Olerup SSP AB

NEWS September 2015

Introducing speed and precision in Real-Time

We are more than SSP A few words from Gordon Hill, General Manager, Olerup Inc. ■ Page 2

XM-ONE® PT-program update This month, the 9th round of the XM-ONE® Proficiency Testing (PT) will be run. ■ Page 3

GAMMA-TYPE™ A new matching technology in stem cell transplantation. ■ Page 6

Olerup real-time PCR kits, QTYPE®, utilize the multiplexing capabilities of hydrolysis probes to increase the capacity for unique assays in each well. Using hydrolysis probes allows robust typing from low-input DNA concentrations, and removes the risk of misinterpreting primer dimers and other PCR artefacts seen in real-time techniques based on intercalating dyes. ■ Page 4

The Olerup Real Time PCR System consists of QTYPE® HLA Typing Kits, the ACCUTYPER® Real Time PCR Instrument and SCORE™6 interpretation software. Together, these three components offer a streamlined workflow for your HLA typing needs. ■ Page 5

Assign™ SBT v471 Introducing analysis of Group Specific Amplification (GSA) data ■ Page 8

Why Olerup should be your top choice for SSP Olerup is dedicated to updating our primer mixes to achieve the best possible kit resolution ■ Page 9

NEWS

We are more than SSP Ever since we pioneered the typing test in the early 1990’s, the name Olerup has been synonymous with SSP. And make no mistake, SSP remains a key area of focus for us…and the alleles keep coming. With a dedicated team, we continue to proactively improve on our SSP foundation by adding new primers and increased multiplexing. For example, since October 2013, when I started with Olerup, we expanded our high resolution DPB1 tray from 48 wells to 87 wells to keep up with the new alleles. We listen more than ever to our customers’ suggestions, to ensure that what we are developing meets and exceeds your needs. This is what sets us apart. Our strength in SSP forms our foundation, but Olerup is more than SSP. After listening to our loyal customers, it occurred to us that we needed to do a better job of communicating what else we do. So, in 2014, we launched Olerup News to highlight our current areas of focus, and to give a glimpse of what’s on the horizon. Olerup provides a comprehensive suite of Sanger SBT kits and software as well as Gamma-Type™, a new matching technology for the gamma block of the MHC. Olerup also provides XM-ONE®, a novel HLA and nonHLA crossmatch test using donor endothelial pre-cursor cells. Most excitingly, we now are positioned to share details of our real time HLA typing assay QTYPE® and qPCR instrument ACCUTYPER®.

New product line – introduction of QTYPE® The innovative Olerup qPCR kits, QTYPE®, will soon be available to our customers in the US and Canada. Olerup is also introducing ACCUTYPER®, a qPCR instrument that will make the laboratory workflow seamless in combination with QTYPE® and SCORE™. SBT – Conexio Partner introduces new kit enhancements: We are pleased to announce the availability of Conexio’s enhanced Class II SBT kits. For the new full exon coverage map, see page 7. Of particular note is a multiplexed DPB1 assay covering Exons 1-5. Worldwide distribution Olerup has more than 25 distributors worldwide; many of them have been supplying customers with Olerup products for more than 15 years. Our distributors stay with us because they know from experience how good Olerup products are and they place great value on the fact that Olerup is the world’s number one SSP supplier. I invite you to visit us at ASHI booth 401 during the conference, and please mark your schedules for Tuesday, Sep 29th at 12:00pm, where we will lead a focused discussion on QTYPE® at a luncheon symposium.

NEWS

Olerup and venues 2015 / 2016 Olerup products and representatives can be seen throughout 2015 and 2016 at the following meetings, conferences and exhibitions. We look forward meeting you.

MEET US AT ASHI September 28-Oct 2, Savannah, Georgia

Arab Health 2016 January 25 – 28, Dubai (UAE)

DGI October 8-10, Lübeck, Germany

EFI - 30th Conference May 11-14, 2016, Kos, Greece

APHIA October 11-14, Bangkok, Thailand

ASHI - 42nd Annual Meeting September 26-30, 2016, St. Louis, MO, USA

Eurotransplant October 15-16, Netherlands MEDICA November 16-19, Düsseldorf, Germany

XM-ONE® PT-program update This month, the 9th round of the XM-ONE® Proficiency Testing (PT) will be run. It is pleasing to see that participation rate is steadily increasing, with the last two rounds counting 19 participating HLA laboratories. We continuously strive to make improvements to the PT program and your input is important for this. Based on participant

Olerup is more than SSP.

PT round

Gordon Hill General Manager, Olerup Inc.

Date

Centers participating

requests, we now also evaluate T and B cell cross-match results for those centers that run XM-ONE® with the 3-in-1 protocol (including CD3 and CD19 detections antibodies for parallel cross matching with EPCs, T cells, and B cells). Based on another request, from 2015 participants can not only report in paper copy form, but we also offer eTables & eReport forms.

Successful participation (for IgG, and IgM if chosen)

2011-I

Sep 2011

5/5

2012-I

Feb 2012

9/10

2012-II

Sep 2012

15/15

2013-I

Mar 2013

14/15

2013-II

Sep 2013

16/17

2014-I

Mar 2014

14/17 *

2014-II

Sep 2014

18/19 **

2015-I

Mar 2015

14/19 ***

* 3/17 requested to be considered ”Educational participants” and were not evaluated. ** 1/19 requested to be considered ”Educational participants” and was not evaluated. *** 3/19 requested to be considered ”Educational participants” and 2 did not report

Are you running XM-ONE® and want to enroll in the PT program? Please contact Eileen Tully, PT-Services Coordinator: eileen.tully@olerup.com, Phone: 877-OLERUP1

XM-ONE® The XM-ONE® transplantation cross match test is today primarily used prior kidney transplantation, to detect non-HLA antibodies against the donor’s endothelia. The results of the first clinical trials using XM-ONE®, published in February 2009 in the medical journal Transplantation, indicated that XM-ONE® could predict rejection reactions in patients that were approved for transplants using test methods that are considered standard. Later publications have shown that XM-ONE® is a valuable complement to traditional antibody testing prior organ transplantation. XM-ONE® is CE marked for sales in Europe and has been cleared by the FDA for sales in the U.S. Read more about XM-ONE® at www.absorber.se.

NEWS

Olerup introduces the QTYPE Real Time PCR System

“By combining hydrolysis probe technology with traditional SSP in a 384 well format we have plenty of scope for adding hundreds of new reactions, meaning we can continually

SCORE™6 Interpretation Software SCORE™6 has been redesigned to improve the workflow with both QTYPE® and SSP products. The workflow has been simplified to just four screens: Patient, Test, Analysis and Report. All of the existing tools and options from SCORE™5 can be found in SCORE™6, and many more. The underlying database technology has been upgraded in SCORE™6, increasing the speed and reliability of the program and allowing multiple users to access the same database at the same time. Users with existing SCORE™6 databases can make use of the migration tool to easily port their data to the new version.

SCORE™6 Workflow

Patient

Test

Analysis

Report

• Enter patient information

• Associate samples with tests

• Review data

• Assemble report

• Enter sample information

• Import data (automatic with ACCUTYPER®)

• Authorise results

• Print or save report

ACCUTYPER® Real Time PCR Instrument The ACCUTYPER® is an affordable yet fully-featured real time PCR instrument from Olerup. The instrument has interchangeable 96 and 384-well blocks, and can excite and detect reactions in up to five channels. The ACCUTYPER® is controlled directly from SCORE™6 when used with QTYPE® kits, but it is also delivered with open research software which can be used for other assays – whether they are probe based, end stage or melt curve. The QTYPE® Real Time PCR Workflow The QTYPE® workflow is simple, whether you have an Olerup-SSP ACCUTYPER® or another real time PCR instrument. Each test is provided with a single use vial of mastermix which contains all of the components necessary for hot start PCR - there is no separate polymerase to add. To set up a test just mix the master mix with your DNA sample and dispense 10 µl into each well of the plate. The plate is then sealed with the included optically-clear sealing sheet and is centrifuged briefly to ensure that there are no bubbles present. The full QTYPE® Real Time PCR system offers the most streamlined workflow. The ACCUTYPER® is controlled directly from SCORE™6, so once a patient and sample is

PREPARE

QTYPE® HLA Typing Kits The QTYPE® HLA Typing Kits are PCR plates with pre-aliquoted primer and probe mixes in each well.

improve the resolution of the kits even with new alignment releases” says Ben Passey at Olerup.

10 min

RUN

The QTYPE® Real Time PCR System consists of QTYPE® HLA Typing Kits, the ACCUTYPER® Real Time PCR Instrument and SCORE™6 interpretation software. Together, these three components offer a streamlined workflow for your HLA typing needs. The QTYPE® HLA Typing Kits and SCORE™6 can also be used in conjunction with the Roche LightCycler 480 II. Other real time PCR instruments, such as or the Life Technologies Viia 7 and QuantStudio 6 & 7 are being validated.

All QTYPE® HLA Typing Kits use hydrolysis probe technology. Like primers, these probes are sequence specific, which means that three cis-located polymorphic regions can be used in determining the specificity of a single reaction. This increases the resolution power of each reaction compared to methods that only use primers. Furthermore, each reaction can be multiplexed using probes with different fluorescent labels. These reactions can be independently read in different detection channels in the real time PCR instrument, meaning each well can contain up to four unique reactions. One channel is reserved for the internal amplification control, leaving three channels for HLA-specific reactions. This gives the potential for 1,152 HLA-specific reactions in a 384 well plate.

50 min ANALYSE

Olerup-SSP is introducing the QTYPE® Real Time PCR System for use in histocompatibility and immunogenetics laboratories that require a rapid system for HLA typing samples from low to intermediate resolution.

10 min

selected, the plate can be inserted into the instrument and the run started from within SCORE™6. When the run is complete the data is automatically transferred from the ACCUTYPER® back to SCORE™6 and interpreted to give the genotype of the sample. For users with other real time PCR instruments, the workflow is also simple. The instrument is controlled from the manufacturer’s software and at the end of the run the data should be exported. SCORE™6 can then import this data and automatically interpret the genotype.

• Prepare DNA/Master Mix • Dispense plate • Seal plate • Vortex and centrifge

• Set up instrument • Insert plate • Run

• Transfer raw data • Perform data analysis • Review results

Ben Passey Development Manager, Olerup SSP AB

NEWS

GAMMA-TYPE – A new matching technology in stem cell transplantation

NEWS

™

Current criteria for selecting unrelated donors in hematopoietic stem cell transplantation (HSCT) include matching of HLA genes within 3 of the four major Major

Histocompatibility Complex (MHC) genomic blocks (i.e. alpha, beta, gamma, and delta).

The gamma block (including the Bf, C2 and C4 genes) has historically been excluded even though this block may provide additional MHC haplotype matching information. Initial studies with the Gamma-Type™ assay support previous study findings that mismatches in the gamma block region may be associated with worse outcome in HSCT.

Conexio’s sequencing based typing kits for HLA Class I and Class II genes, covering all the required NMDP null alleles. Data fully compatible with Assign™ SBT sequence analysis software that includes analysis of non-coding regions. SBT Resolver™ Amplicon Coverage map 5’UTR Exon Intron Exon Intron Exon Intron 1 1 2 2 3 3

Exon Intron Exon Intron Exon Intron Exon Intron Exon 4 4 5 5 6 6 7 7 8

At the BMT tandem meeting in San Diego, Feb 11-15, 2015, Dr. Medhat Askar and colleagues at the Cleveland Clinic presented initial findings with Gamma-Type™. Investigating 236 HSCT recipient/unrelated donor pairs transplanted at their center (20002010), they concluded that specific gamma block mismatches are associated with a higher risk of severe acute GVHD, independent of the HLA and MICA match and other clinical risk factors. Larger, retrospective, studies including more than 700 donor-patient pairs are currently underway at centers in the US.

Class II updates:

Two different kit formats 25-test kit; KD-PD8.0-1(25) Suitable for patient-donor testing. Contains pre-aliquoted trays each with PCR-SSP mixes for the typing of 25 gamma block specific SNPs. Sufficient for 25 DNA samples. 96-test kit; KD-PD8.0-1(96 Suitable for retrospective studies. Bulk format, with each kit containing 25 different vials of PCR mixes for the typing of 25 unique gamma block specific SNPs. One kit is sufficient for 96 DNA samples.

: Complete sequencing of all exon 3 allele groups in addition to exon 2 : Complete coverage of exon 2 and exon 3 : Complete coverage of exon 1, 3, 4 and 5 in addition to exon 2 Distributed by For Research Use Only. Not for use in diagnostic procedures.

No claim or representation is intended to provide information for the diagnosis, prevention, or treatment of a disease.

www.olerup.com

Designed, developed and manufactured by Conexio Gamma-Type™ is for Research Use Only. Not for use in diagnostic procedures. No claim or representation is intended to provide information for the diagnosis, prevention or treatment of a disease.

conexio-genomics.com

Pioneering since 2002

NEWS

Assign™ SBT v471 – Introducing analysis of Group Specific Amplification (GSA) data Conexio Genomics continuously works to improve the Assign™ SBT software. Assign™ SBT v471 is the latest version of the renowned SBT analysis software. This new release features improvements to the display, an Auto Save function and full HARPSFinder™ functionality for prospective HARPS® selection. The new HARPSFinder™ function replaces the on-line version. As the previous version (Assign™ SBT v4.7), the new release is fully

compatible with Conexio´s SBT Resolver™ typing kits and HARPS®. The software can also be used standalone with alternative sequencing based typing approaches.

(GSA) methods. Furthermore, data combinations of locus specific amplification, HARPS® and GSA can be also be analysed.

Assign™ SBT v471 is extremely flexible as it allows not only analysis of data generated with Locus Specific Amplification (LSA) methods, but can also be configured for analysis of data produced by Group Specific Amplification

NEWS

Why Olerup should be your top choice for SSP Olerup is dedicated to updating our primer mixes to achieve the best possible kit resolution Olerup SSP AB does not just supply the same SSP kits year after year. We provide SSP kits that are up-to-date with the recently released allele database at the time of batch manufacturing. Up-to-date kits, of course, means more than just updating kit documentation. It is about updating the actual primer mixes to improve resolution of the kit to address newly assigned alleles. This approach is the Olerup

approach. It demands continuous investments into the product line, a dedicated production lab, as well as significant pre-production primer mix optimization. Additionally, updating kits for new alleles is not only adding new wells and expanding the kit formats. Much work is put into multiplexing in order to keep the product formats

as slim as possible. Table 1 below reflects our recent work in this area. New batches that were produced during the first half of 2015 included 2621 unique primer mixes. Of these, 22% were new designs, either through multiplexing within previously existing designs (19.6%) or through adding new wells to the kits (2.4%).

Table 1. Outlines the number of primer mixes (wells) that have been produced and updated for new alleles across the complete Olerup SSP product line during the first six months of 2015.

Olerup SSP® allele updating work in numbers across the product line, Jan-July 2015 Total number of produced and unique primer mixes (wells)

2621

Out of which: Multiplexing instances for resolution of new alleles

513

Added wells

Sum of changes on primer mix level for allele updating

577

We take great pride at Olerup SSP AB in our continuous updating of the primer mixes prior to batch production. This allows us to provide SSP trays that are up-to-date in the truest sense. Over the years, the increasing number of assigned alleles has generated many challenges to the manufacturers of HLA typing assays. We have adopted strategies to address these challenges and are straightforward in providing customers with information about our improvements and changes.

Assign™ SBT v471 automatically combines GSA data. The GSA 1 and GSA 2 layers in top screenshot represent a first set and a second set of GSA data for the sample. MM 0 layer represents the heterozygous consensus for the sample (both GSA layers combined). Assign™ SBT v471 is for Research Use Only. Not for use in diagnostic procedures. No claim or representation is intended to provide information for the diagnosis, prevention or treatment of a disease.

as an HLA allele that has been sequenced by more than a single laboratory or from multiple sources. This status is updated for each new database. We consider all nulls, as well as polymorphisms outside of the region encoding the peptide binding domain. Currently, the HLA-A, B, C, DRB1, DQB1 and DPB1 high resolution kits are updated according to this strategy prior to each batch production.

The Olerup SSP® Add-ons - Time saving solutions to meet requirements As a result of new null allele requirements in 2015 by NMDP for unrelated HSCT, Olerup SSP AB has just recently expanded the “Add-on” line to also include kits specifically targeted these nulls. The result is a product portfolio of small SSP kits for smart solutions covering all null allele testing required by the NMDP.

Flexible solutions for SSP low resolution typing In 2012, Olerup SSP AB launched the expanded versions of the single locus low resolution trays, giving our customers the choice between the classical screening/combi formats and the expanded single-locus formats with increased resolution. Now in 2015 we can confirm that our decision to move in this direction was a good one. The outcome is increasingly popular low resolution kits with more redundancy, greater resolution, and increased resistance to ambiguities. A high resolution line which keeps up The updating strategy for our high res kits has also changed over the years. To be able to accommodate the large sets of newly assigned alleles, Olerup has decided to focus high resolution kit updating on confirmed alleles, which are defined by the IMGT/HLA database

Anna Hedlund, Head of R & D at Olerup SSP

NEWS

Olerup - Global distribution to HLA laboratories At Olerup, we strive to continuously develop and improve our web services. We believe that information and news of our products and companies should be easy accessible for users. If you have not recently visited our web pages, please do so and see how they can be of value for you. Olerup web pages are easiest to access through www.olerup.com, the portal to our

distribution network. Among other information, you can find news about our companies, see what meetings Olerup will attend and, of course, get further information about our different products.

has been working with us for many years and are well-established partners to HLA laboratories in their respective region/country. Nevertheless, every year we add new distributors to our global network.

Distributor contact information is also easily accessible at the web page. Olerup is currently represented in more than 50 countries around the world. The majority of our distributors

A Tool for Enhanced Risk Assessment

Improving outcomes in kidney transplantation: Non-HLA antibodies and Enhanced Clinical Risk Assessment The XM-ONE® transplantation cross match is used prior kidney transplantation, to detect non-HLA antibodies against the donor’s endothelia. Results from the first clinical trial using XM-ONE®, published in February 2009 in the medical journal Transplantation1, demonstrated that XM-ONE® could predict rejection reactions in patients that were approved for transplants using test methods that are considered standard. Later publications have shown that XM-ONE® is a valuable complement to traditional antibody testing prior organ transplantation.

GERMANY/AUSTRALIA/BENELUX

Olerup Inc Orders and support: info.us@olerup.com

Olerup GmbH Orders and support: support-at@olerup.com

Tel: +1-877-OLERUP1 Fax: +1-610-344-7989

Tel: +43-1-710 15 00 00 Fax: +43-1-710 15 00 10

901 S. Bolmar St., Suite R West Chester, PA 19382

Lowengasse 47/6 1030 Vienna, Austria

NORDIC (DENMARK, FINLAND, ICELAND, NORWAY, SWEDEN AND THE BALTICS)

OTHER REGIONS

XM-ONE® is CE marked for sales in Europe and has been cleared by the FDA for sales in the U.S. Read more about XM-ONE® at www.absorber.se Breimer, M, Rydberg, L, Jackson, AM et al, Multicenter evaluation of a novel endothelial cell crossmatch test in kidney transplantation, Transplantation 2009, 87(4):549-556.

To place your order please use the Nordic Web Shop or contact our sales team through info@olerup.com

Olerup product lines are in other regions made available through our network of specialized distributors. To find your distributor please go to http://www.olerup.com/distributors/

Tel: +46-8-50893900 Fax: +46-8-7178818 Franzéngatan 5 112 51 Stockholm, Sweden

Manufactured by:

Distributed by:

www.absorber.se

www.olerup.com

OLE-150921A-1

NEWS

Introducing QTYPE

Speed and Precision in Real Time By combining the multiplexing power, sensitivity and specificity of hydrolysis probes, QTYPE® is a quick, robust, and easy-to-use alternative in HLA typing. The technology ensures improved resolution whilst retaining plenty of capacity to cope with future allele updates. As part of our real time concept, we also provide ACCUTYPER®, a reliable and convenient qPCR- instrument that will make the workflow with QTYPE® seamless. Data handling and result interpretation is of course powered by SCORE™

QT-150908 A-1

For Research Use Only.

Manufactured by:

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