AzTrials
AzTrials

Kidney Transplant RejectionNCT04239703

Trifecta-Kidney cfDNA-MMDx Study

An observational study of Kidney Transplant Rejection, run by University of Alberta at 31 sites in 10 countries.

Recruiting

Looking for participants now. Contact the study team or a site below to ask about taking part.

Sites and contacts
Official record
Search all trials

Trial summary

Demonstrate the relationship between DD-cfDNA levels and HLA antibodies in blood, and the Molecular Microscope® (MMDx) Diagnostic System results in indication biopsies. Read more about this study

Who is it for?
Any age
Conditions the study is for
Kidney Transplant Rejection
What is being tested?
MMDx, Prospera and HLA antibody
How many people
300 participants
Where
31 sitesin 10 countries
Study dates
1 Dec 2019to Dec 2029
Phase
Observational
Are there results?
Not yet: the study is under way

Key requirements

Who can take part, as listed by the sponsor. Only the study team can confirm whether you are eligible, so take this list to your doctor.

  • Ages Any age
  • All sexes
  • No healthy volunteers
  • Child, Adult, Older adult

Study population: The study population includes patients with a functioning kidney transplant undergoing a biopsy for clinical indications as standard of care to determine the cause of their graft dysfunction (deterioration in graft function, delayed graft function, proteinuria).

Who may be able to join

  • All kidney transplant recipients undergoing a kidney biopsy for clinical indications, as determined by their physician or surgeon, will be eligible to enroll in the study.

Who cannot join

  • Patients will be excluded from the study if they decline participation or are unable to give informed consent or multiple organ recipients.

Trial locations and contacts

31 sites in 10 countries. Contact the study team or a site near you to ask about taking part.

Konrad S Famulski, PhD contact

1 780 492 1725

konrad@ualberta.ca

Robert Polakowski, PhD contact

1 780 492 5091

polakows@ualberta.ca

31 sites in 10 countries

United States11 sites, 11 recruiting

  • University of Maryland School of Medicine

    Baltimore, Maryland, 21201

    Raissa Toure, RToure@som.umaryland.edu

    Jonathan Bromberg, MD PhD

    Recruiting

  • The Johns Hopkins University, School of Medicine

    Baltimore, Maryland, 21205

    Darin B Ostrander, PhD, 410-614-6702, dostran1@jhmi.edu

    Daniel C Brennan, MD

    Recruiting

  • University Hospitals Cleveland Medical Ctr.

    Cleveland, Ohio, 44106-5048

    Katherine R Carter, Katherine.carter@uhhospitals.org

    Jittirat Arksarapuk, MD

    Recruiting

  • Cleveland Clinic

    Cleveland, Ohio, 44195

    Debra Camino, caminod@ccf.org

    Emillio Poggio, MD

    Recruiting

  • Detroit Medical Center, Harper University Hospital of Wayne State University

    Detroit, Michigan, 48201

    Rajeev Sharma, MD, rasharma@med.wayne.edu

    Rajeev Sharma, MD

    Not yet recruiting

  • Henry Ford Hospital

    Detroit, Michigan, 48202

    Iman Francis, Ifranci1@hfhs.org

    Milagros Samaniego-Picota, MD

    Recruiting

  • Intermountain Transplant Services

    Murray, Utah, 84107

    Jake Krong, Jake.Krong@imail.org

    Sanjiv Anand, MD

    Recruiting

  • Virginia Commonwealth University Medical Center

    Richmond, Virginia, 23298

    Gaurav Gupta, MD, ggupta@mcvh-vcu.edu

    Gaurav Gupta, MD

    Recruiting

  • Division of Nephrology & UW Organ Transplant Center University of Washington

    Seattle, Washington, 98195

    Chris Blosser, MD, CBlosser@nephrology.washington.edu

    Chris Bosser, MD

    Recruiting

  • Barnes-Jewish Hospital, Washington University at St. Louis

    St Louis, Missouri, 63110

    Andrew Malone, amalone@wustl.edu

    Andrew Malone, MD PhD

    Recruiting

  • Tampa General Hospital

    Tampa, Florida, 33606

    Natalie Remsen, nremsen@tgh.org

    Rajendra Baliga, MD

    Recruiting

Other trials in United States

Poland11 sites, 8 recruiting

  • Department of Nephrology and Transplantation Medical University in Bialystok

    Bialystok, 15-540

    Completed

  • University Hospital nr1 Bydgoszcz, Klinika Transplantologii

    Bydgoszcz, 85-094

    Completed

  • Medical University of Gdańsk Klinika Nefrologii Transplantologii i Chorób Wewnętrznych

    Gdansk, 80-259

    Andrzej Chamienia, MD, chamien@gumed.edu.pl

    Alicja Dębska-Ślizień, MD

    Recruiting

  • Medical University of Silesia

    Katowice, 40-027

    Grzegorz Piecha, MD, g.piecha@outlook.com

    Grzegorz Piecha, MD

    Recruiting

  • Department of Transplantation and General Surgery, Wojewodzki Hospital

    Poznan, 60-479

    Maciej Glyda, MD, glydam@wp.pl

    Maciej Glyda, MD

    Recruiting

  • Department of Nephrology, Transplantation and Internal Medicine, University Hospital n.2

    Szczecin, 70-111

    Leszek Domański, MD, domanle@pum.edu.pl

    Leszek Domański, MD

    Recruiting

  • Pomeranian Medical University, Samodzielny Publiczny Woj. Szpital Zespolony, Oddzial Nefrologii i Transplantacji Nerek

    Szczecin, 71-455

    Marek Myślak, MD Phd, marek.myslak@pum.edu.pl

    Marek Myślak, MD PhD

    Recruiting

  • Medical University of Warsaw, Department of Transplantation Medicine, Nephrology and Internal Diseases

    Warsaw, 02-006

    Magdalena Durlik, MD, magdalena.durlik@wum.edu.pl

    Magdalena Durlik, MD

    Recruiting

  • Transplant Medicine & Nephrology Clinic, Medical University of Warsaw

    Warsaw, 02-006

    Agnieszka Perkowska-Ptasińska, MD PhD, aggape@poczta.onet.pl

    Agnieszka Perkowska-Ptasińska, MD PhD

    Recruiting

  • The Children's Memorial Health Institute, Department of Nephrology, Kidney Transplantation and Hypertension

    Warsaw, 04-730

    Completed

  • Wroclaw Medical University, Department of Nephrology and Transplantation Medicine

    Wroclaw, 50-556

    Mirosław Banasik, MD, m.banasik@interia.pl

    Mirosław Banasik, MD

    Recruiting

Other trials in Poland

Canada2 sites, 2 recruiting

  • University of Alberta, Department of Medicine

    Edmonton, Alberta, T6G 2G3

    Soroush Shojai, MD, shojai@ualberta.ca

    Soroush Shojai, MD

    Recruiting

  • ST. Paul's Hospital, 6A Providence Building, 1081 Burrard Street

    Vancouver, British Columbia, V6Z 1Y6

    Angela Ogniben, AOgniben@providencehealth.bc.ca

    John S Gill, MD, MS

    Recruiting

Other trials in Canada

Australia1 site, 1 recruiting

  • Department of Nephrology, The Royal Melbourne Hospital 1 South East

    Melbourne, VIC 3050

    Peter D Hughes, MD, peter.hughes@mh.org.au

    Kevin Chow, MD

    Recruiting

Other trials in Australia

Croatia1 site, 1 recruiting

  • University Hospital Merkur Renal Division

    Zagreb, 1910000

    Zeljka Jurekovic, MD, zeljka.jurekovic@gmail.com

    Mladen Knotek, MD

    Recruiting

Other trials in Croatia

Czechia1 site, 1 recruiting

  • Institute for Clinical and Experimental Medicine (IKEM)

    Prague, 140 21 Praha 4

    Petra Hruba, MD, hrup@ikem.cz

    Ondrej Viklicky, MD

    Recruiting

Other trials in Czechia

Germany1 site, 1 recruiting

  • Charite-Medical University of Berlin Department of Nephrology

    Berlin, 10117

    Klemens Budde, MD, klemens.budde@charite.de

    Monique Greiner-Pol, monique.greiner-pol@charite.de

    Recruiting

Other trials in Germany

Lithuania1 site, 1 recruiting

  • Centre of Nephrology, Vilnius University Hospital Santaros Klinikos

    Vilnius, LT-08661

    Alvita Vickiene, alvita.gincaite@gmail.com

    Marius Miglinas, MD

    Recruiting

Other trials in Lithuania

Slovenia1 site, 1 recruiting

  • Department of Nephrology, University Medical Centre

    Ljubljana, 21000

    Nika Kojc, MD, nika.kojc@mf.uni-lj.si

    Miha Arnol, MD

    Recruiting

Other trials in Slovenia

Switzerland1 site

  • University Hospital Zurich

    Zurich, 8091

    Completed

Other trials in Switzerland

About this study

Official title: Trifecta-Kidney cfDNA-MMDx Study: Comparing the DD-cfDNA Test to MMDx Microarray Test, Central HLA Antibody Test, and Histology.

Demonstrate the relationship between DD-cfDNA levels and HLA antibodies in blood, and the Molecular Microscope® (MMDx) Diagnostic System results in indication biopsies.

Read the detailed description

There is a need for better screening of kidney transplant patients for rejection. Patients with kidney transplants are routinely tested (creatinine, urine protein, histology and donor specific antibody (DSA) as standard of care to detect rejection, but these tests are not adequate. Rejection is often missed by these tests (false negatives) and other processes such as acute kidney injury can produce false-positive results. Moreover, histology has a high interobserver disagreement diagnosing rejection, and cannot accurately assess acute injury. A definitive molecular assessment of rejection and injury in kidney biopsies has emerged - the Molecular Microscope® Diagnostic System (MMDx) - developed by the Alberta Transplant Applied Genomics Centre, University of Alberta. Now a new screening test is being introduced: the monitoring of donor-derived cell-free DNA (DD-cfDNA) released in the blood by the kidney during rejection. The Natera Inc DD-cfDNA Prospera® test is based on the massively multiplex PCR that targets 13,392 single nucleotide polymorphisms and targeted sequences are quantified by Next Generation Sequencing. The Prospera® test done on kidney transplant recipients detected "active rejection" and differentiated it from borderline rejection and no rejection. It is likely, however, that DD-cfDNA test may miss some T cell-mediated rejection (TCMR) cases and the distinction between early and fully developed antibody-mediated rejection (ABMR) was not tested. No study has actually examined the DD-cfDNA results in kidney transplants with acute or chronic kidney disease (AKI and CKD). DD-cfDNA measurements have only been correlated with histology, a flawed standard. DD-cf-DNA test must now be calibrated against MMDx that is based on global gene expression, the new standard for biopsy interpretation. The present study will calibrate centrally measured (Natera Inc) DD-cfDNA levels obtained at the time of an indication biopsy against the MMDx measurements of TCMR, and ABMR (early-stage, fully-developed, and late-stage), AK, and atrophy-fibrosis. We will compare blood DD-cfDNA measurements in 600 samples at the time of 300 indication biopsies to the MMDx results, as well as central assessment of HLA antibody (One Lambda) in 300 blood samples, interpreted centrally as DSA based on the tissue typing results. This study is an extension of the INTERCOMEX ClinicalTrials.gov Identifier: NCT01299168. Investigators have collected 1195 kidney biopsies, 1103 blood samples for DD-cfDNA test and 1150 blood samples for One Lamba test, and will extend this study to the total of 1400 biopsies and 2800 blood samples.

Being tested
MMDx, Prospera, HLA antibody
Keywords
donor derived cell-free DNA, blood, kidney biopsy

What participants receive

What participants receive. In a randomised study each person is assigned to one group by chance.

  • MMDxDiagnostic test

    Portion of kidney transplant indication biopsy

  • ProsperaDiagnostic test

    Also called transplant patient blood sample

    Transplant patient blood sample

  • HLA antibodyDiagnostic test

    Also called transplant patient blood sample

    Transplant patient blood sample

Study groups in NCT04239703
Study groupRoleWhat they receive
Kidney transplant biopsies for causeThe study population includes patients with a functioning kidney transplant undergoing a biopsy for clinical indications as standard of care.
Receives: Diagnostic Test: MMDx; Diagnostic Test: Prospera; Diagnostic Test: HLA antibody

What is being measured

The primary outcome is the main question the study is designed to answer.

  1. Calibration of Prospera test for T cell-mediated rejection

    Calibration of DD-cfDNA test cut-off values against the probability of T cell-mediated rejection in the biopsy as reported by MMDx.

    Measured: 18 months

  2. Calibration of Prospera test for antibody-mediated rejection

    Calibration of DD-cfDNA test cut-off values against the probability of antibody-mediated rejection in the biopsy as reported by MMDx.

    Measured: 18 months

  3. Calibration of Prospera test for kidney injury

    Calibration of DD-cfDNA test cut-off values against the probability of acute and chronic kidney injury in the biopsy as reported by MMDx.

    Measured: 18 months

  4. Report calibrated Prospera test results for rejection

    Report new DD-cfDNA test cut-off values for rejection

    Measured: 6 months

  5. Report calibrated Prospera test results for kidney injury

    Report new DD-cfDNA test cut-off values for acute and chronic kidney injury

    Measured: 6 month

Secondary outcomes (2)

  1. Determine if Prospera blood test can replace kidney biopsy test

    Determine if Prospera test, as calibrated by this DD-cfDNA-HLA-MMDx study, will avoid need for indication biopsy when kidney transplant function deteriorates. This will be based on the consensus between participating clinicians.

    Measured: 6 months

  2. Assessment of donor-specific antibody status

    Report and compare the DSA status based on centralized and local HLA antibody measurement.

    Measured: 6 months

Timeline

From the start of the study to its completion; the mark is today. Estimated dates are the sponsor’s plan.

  1. First posted27 Jan 2020
  2. Study start1 Dec 2019
  3. Primary completionDec 2028 (estimated)
  4. Study completionDec 2029 (estimated)

Resources

More details

For doctors and researchers: how the study is designed, who runs it, and its registry IDs.

Study design
Study type
Observational
Observational model
Cohort
Time perspective
Prospective
Enrolment
300 participants (estimated)
IDs, dates and publications
ClinicalTrials.gov ID
NCT04239703
Sponsor's protocol ID
ATAGC05
Status last verified
Jun 2026
First submitted
14 Jan 2020
Last updated
2 Jun 2026
Results posted
No
FDA-regulated drug
No
FDA-regulated device
No
Individual data sharing
No

Publications

  • Madill-Thomsen KS, Halloran PF. Precision diagnostics in transplanted organs using microarray-assessed gene expression: concepts and technical methods of the Molecular Microscope(R) Diagnostic System (MMDx). Clin Sci (Lond). 2024 Jun 5;138(11):663-685. doi: 10.1042/CS20220530.

    PubMed 38819301

  • Halloran PF, Madill-Thomsen KS. Donor-derived Cell-free DNA: A Step Forward in the Quest for Transplant Truth. Transplantation. 2025 Jun 1;109(6):910-914. doi: 10.1097/TP.0000000000005332. Epub 2025 Jan 28. No abstract available.

    PubMed 39883025

  • Halloran PF, Reeve J, Madill-Thomsen KS, Demko Z, Prewett A, Billings P; Trifecta Investigators. The Trifecta Study: Comparing Plasma Levels of Donor-derived Cell-Free DNA with the Molecular Phenotype of Kidney Transplant Biopsies. J Am Soc Nephrol. 2022 Feb;33(2):387-400. doi: 10.1681/ASN.2021091191. Epub 2022 Jan 20.

    PubMed 35058354

  • Halloran PF, Reeve J, Madill-Thomsen KS, Demko Z, Prewett A, Gauthier P, Billings P, Lawrence C, Lowe D, Hidalgo LG; the Trifecta Investigators. Antibody-mediated Rejection Without Detectable Donor-specific Antibody Releases Donor-derived Cell-free DNA: Results From the Trifecta Study. Transplantation. 2023 Mar 1;107(3):709-719. doi: 10.1097/TP.0000000000004324. Epub 2023 Feb 21.

    PubMed 36190186

  • Halloran PF, Madill-Thomsen KS, Reeve J. The Molecular Phenotype of Kidney Transplants: Insights From the MMDx Project. Transplantation. 2024 Jan 1;108(1):45-71. doi: 10.1097/TP.0000000000004624. Epub 2023 Dec 13.

    PubMed 37310258

  • Halloran PF, Reeve J, Madill-Thomsen KS, Kaur N, Ahmed E, Cantos C, Al Haj Baddar N, Demko Z, Liang N, Swenerton RK, Zimmermann BG, Van Hummelen P, Prewett A, Rabinowitz M, Tabriziani H, Gauthier P, Billings P; Trifecta Investigators*. Combining Donor-derived Cell-free DNA Fraction and Quantity to Detect Kidney Transplant Rejection Using Molecular Diagnoses and Histology as Confirmation. Transplantation. 2022 Dec 1;106(12):2435-2442. doi: 10.1097/TP.0000000000004212. Epub 2022 Nov 22.

    PubMed 35765145

  • Gauthier PT, Madill-Thomsen KS, Demko Z, Prewett A, Gauthier P, Halloran PF; Trifecta-Kidney Investigators. Distinct Molecular Processes Mediate Donor-derived Cell-free DNA Release From Kidney Transplants in Different Disease States. Transplantation. 2024 Apr 1;108(4):898-910. doi: 10.1097/TP.0000000000004877. Epub 2023 Dec 27.

    PubMed 38150492

  • Hidalgo LG, Madill-Thomsen KS, Reeve J, Mackova M, Gauthier P, Demko Z, Prewett A, Lee M, Alhamad T, Anand S, Arnol M, Baliga R, Banasik M, Blosser CD, Bobba S, Brennan D, Bromberg J, Budde K, Chamienia A, Chow K, Ciszek M, Costa N, Deborska-Materkowska D, Debska-Slizien A, Domanski L, Fatica R, Francis I, Fryc J, Gill J, Gill J, Glyda M, Gourishankar S, Gryczman M, Gupta G, Hruba P, Hughes P, Jittirat A, Jurekovic Z, Kamal L, Kamel M, Kant S, Kojc N, Konopa J, Kumar D, Lan J, Lowe D, Mazurkiewicz J, Miglinas M, Moinuddin I, Mueller T, Myslak M, Naumnik B, Paczek L, Patel A, Perkowska-Ptasinska A, Piecha G, Poggio E, Bloudickova SR, Regele H, Schachtner T, Shojai S, Sikosana MLN, Slatinska J, Smykal-Jankowiak K, Haler ZV, Viklicky O, Vucur K, Weir MR, Wiecek A, Zaky Z, Halloran PF. Improving the histologic detection of donor-specific antibody-negative antibody-mediated rejection in kidney transplants. Am J Transplant. 2026 Jan;26(1):117-130. doi: 10.1016/j.ajt.2025.08.029. Epub 2025 Aug 23.

    PubMed 40854490

  • Madill-Thomsen KS, Hidalgo LG, Demko ZP, Gauthier PM, Prewett A, Lowe D, Chang JJ, Mackova M, Budde K, Bromberg JS, Halloran PF; Trifecta-Kidney Study Group. Defining Relationships Among Tests for Kidney Transplant Antibody-Mediated Rejection. Kidney Int Rep. 2025 Jun 17;10(9):3225-3238. doi: 10.1016/j.ekir.2025.06.017. eCollection 2025 Sep.

    PubMed 40980666

  • Halloran PF, Chang J, Mackova M, Madill-Thomsen KS, Akalin E, Alhamad T, Anand S, Arnol M, Baliga R, Banasik M, Blosser CD, Bohmig G, Brennan D, Bromberg J, Budde K, Chamienia A, Chow K, Ciszek M, de Freitas D, Deborska-Materkowska D, Debska-Slizien A, Djamali A, Domanski L, Durlik M, Einecke G, Eskandary F, Fatica R, Francis I, Fryc J, Gill J, Gill J, Glyda M, Gourishankar S, Gryczman M, Gupta G, Hruba P, Hughes P, Jittirat A, Jurekovic Z, Kamal L, Kamel M, Kant S, Kojc N, Konopa J, Lan J, Mannon RB, Matas A, Mazurkiewicz J, Miglinas M, Mueller T, Myslak M, Narins S, Naumnik B, Patel A, Perkowska-Ptasinska A, Picton M, Piecha G, Poggio E, Rajnochova Bloudickova S, Schachtner T, Shojai S, Sikosana ML, Slatinska J, Smykal-Jankowiak K, Solanki A, Veceric Haler Z, Viklicky O, Vucur K, Weir MR, Wiecek A, Wlodarczyk Z, Yang H, Zaky Z, Gauthier PT, Hinze C. A cross-sectional study of the role of epithelial cell injury in kidney transplant outcomes. JCI Insight. 2025 Apr 15;10(10):e188658. doi: 10.1172/jci.insight.188658. eCollection 2025 May 22.

    PubMed 40232852

  • Halloran PF, Reeve J, Mackova M, Madill-Thomsen KS, Demko Z, Olymbios M, Campbell P, Melenovsky V, Gong T, Hall S, Stehlik J. Comparing Plasma Donor-derived Cell-free DNA to Gene Expression in Endomyocardial Biopsies in the Trifecta-Heart Study. Transplantation. 2024 Sep 1;108(9):1931-1942. doi: 10.1097/TP.0000000000004986. Epub 2024 Aug 20.

    PubMed 38538559

  • Halloran PF, Madill-Thomsen KS. The Molecular Microscope Diagnostic System: Assessment of Rejection and Injury in Heart Transplant Biopsies. Transplantation. 2023 Jan 1;107(1):27-44. doi: 10.1097/TP.0000000000004323. Epub 2022 Dec 8.

    PubMed 36508644

Questions about this trial

What is this clinical trial studying?

An observational study of Kidney Transplant Rejection, run by University of Alberta at 31 sites in 10 countries.

The full registered description is in the About this study section and the official record.

Who may be eligible to take part?

The sponsor lists 1 inclusion and 1 exclusion criteria. Eligibility is decided by the study team after screening, so read the full criteria and contact a listed site.

Is this clinical trial currently recruiting?

The record says: looking for participants now, as of the sponsor's last update on 2 Jun 2026. Status can change between updates, so confirm with the study contact before making plans.

Where is the study taking place?

The record lists 31 sites in United States, Poland, Canada, Australia, Croatia and 5 more countries. Whether a site is taking part right now should be confirmed with that site. See every site.

What are the possible benefits and risks?

They depend on the study and the treatment, and no study can promise a benefit. The study team explains the known risks during informed consent, before anyone joins; the record itself does not list them.

Are results available for this study?

This study is still under way. Results are not available in the source record yet.

How can a doctor refer a patient or contact the research team?

Contact Konrad S Famulski, PhD (1 780 492 1725, konrad@ualberta.ca), or a site near the patient. AzTrials does not enrol participants or take referrals.

What does the primary outcome measure?

Calibration of Prospera test for T cell-mediated rejection (time frame: 18 months). The study lists 5 primary outcomes in all. It is what the study was designed to measure, not a finding.

Where can I read the original study information?

On the ClinicalTrials.gov record for NCT04239703, which also lists 12 linked publications.

Does a listing mean the treatment is approved or proven to work?

No. A listing means the study is registered. It does not show that the treatment is safe, works, or is approved by a regulator.

How often is this information updated?

AzTrials copies changed records from ClinicalTrials.gov every night and reloads the whole registry every week. The sponsor last updated this record on 2 Jun 2026.

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