Bérangère Joly1, Alain Stepanian2, David Hajage3, Sandrine Thouzeau4, Sophie Capdenat1, Paul Coppo5, Agnès Veyradier6. 1. Service d'Hématologie biologique, Hôpital Antoine Béclère, Assistance Publique-Hôpitaux de Paris, Clamart, Université Paris 11, France. 2. Service d'Hématologie biologique, Hôpital Louis Mourier, Assistance Publique-Hôpitaux de Paris, Colombes, Université Paris 7, France. 3. Département d'épidémiologie et de recherche clinique, Hôpital Louis Mourier, Assistance Publique-Hôpitaux de Paris, Colombes, Université Paris 7, France. 4. Service d'Hématologie biologique, Hôpital Antoine Béclère, Assistance Publique-Hôpitaux de Paris, Clamart, Université Paris 11, France; French Reference Center for Thrombotic Microangiopathies (CNR-MAT), Hôpital Saint Antoine, Assistance Publique-Hôpitaux de Paris, Paris, Université Paris 6, France. 5. French Reference Center for Thrombotic Microangiopathies (CNR-MAT), Hôpital Saint Antoine, Assistance Publique-Hôpitaux de Paris, Paris, Université Paris 6, France; Service d'Hématologie, Hôpital Saint Antoine, Assistance Publique-Hôpitaux de Paris, Paris, France. 6. Service d'Hématologie biologique, Hôpital Antoine Béclère, Assistance Publique-Hôpitaux de Paris, Clamart, Université Paris 11, France; French Reference Center for Thrombotic Microangiopathies (CNR-MAT), Hôpital Saint Antoine, Assistance Publique-Hôpitaux de Paris, Paris, Université Paris 6, France. Electronic address: agnes.veyradier@abc.aphp.fr.
Abstract
INTRODUCTION: Thrombotic thrombocytopenic purpura (TTP) is a thrombotic microangiopathy (TMA), related to a severe functional deficiency of ADAMTS13 activity (<10% of normal). ADAMTS13 activity is thus crucial to confirm the clinical suspicion of TTP, to distinguish it from other TMAs, and to perform the follow-up of TTP patients. MATERIAL AND METHODS: We compared the performance of the commercial chromogenic assay Technozym ADAMTS13 Activity ELISA (chromogenic VWF73 substrate, Chr-VWF73, Technoclone, Vienna, Austria), to that of our in-house FRETS-VWF73 used as reference method. A large group of 247 subjects (30 healthy volunteers and 217 patients with miscellaneaous TMAs) was studied. RESULTS: The lower limit of detection of the Chr-VWF73 was 3%, which is well adapted to the clinically relevant threshold for TTP diagnosis (10%). Our results showed a reasonable agreement between FRETS-VWF73 and Chr-VWF73 assays to distinguish samples with an ADAMTS13 activity <10% from those with an ADAMTS13 activity >10%. However, Chr-VWF73 assay provided false negative results in ~12% of acute TTP patients. Inversely, the Chr-VWF73 assay globally underestimated ADAMTS13 activity in detectable values ranging from 11 to 100% (with a great variability compared to FRETS-VWF73), which may be a concern for the follow-up of TTP patients in remission. CONCLUSION: In-house assays developed and performed by expert laboratories remain the reference methods that should be used without limitation to control values provided by commercial assays when needed. Also, the development of an international reference preparation will be crucial to improve standardization.
INTRODUCTION:Thrombotic thrombocytopenic purpura (TTP) is a thrombotic microangiopathy (TMA), related to a severe functional deficiency of ADAMTS13 activity (<10% of normal). ADAMTS13 activity is thus crucial to confirm the clinical suspicion of TTP, to distinguish it from other TMAs, and to perform the follow-up of TTP patients. MATERIAL AND METHODS: We compared the performance of the commercial chromogenic assay Technozym ADAMTS13 Activity ELISA (chromogenic VWF73 substrate, Chr-VWF73, Technoclone, Vienna, Austria), to that of our in-house FRETS-VWF73 used as reference method. A large group of 247 subjects (30 healthy volunteers and 217 patients with miscellaneaous TMAs) was studied. RESULTS: The lower limit of detection of the Chr-VWF73 was 3%, which is well adapted to the clinically relevant threshold for TTP diagnosis (10%). Our results showed a reasonable agreement between FRETS-VWF73 and Chr-VWF73 assays to distinguish samples with an ADAMTS13 activity <10% from those with an ADAMTS13 activity >10%. However, Chr-VWF73 assay provided false negative results in ~12% of acute TTP patients. Inversely, the Chr-VWF73 assay globally underestimated ADAMTS13 activity in detectable values ranging from 11 to 100% (with a great variability compared to FRETS-VWF73), which may be a concern for the follow-up of TTP patients in remission. CONCLUSION: In-house assays developed and performed by expert laboratories remain the reference methods that should be used without limitation to control values provided by commercial assays when needed. Also, the development of an international reference preparation will be crucial to improve standardization.
Authors: Marie Scully; Paul Knöbl; Karim Kentouche; Lawrence Rice; Jerzy Windyga; Reinhard Schneppenheim; Johanna A Kremer Hovinga; Michiko Kajiwara; Yoshihiro Fujimura; Caterina Maggiore; Jennifer Doralt; Christopher Hibbard; Leah Martell; Bruce Ewenstein Journal: Blood Date: 2017-09-14 Impact factor: 22.113
Authors: Nicolas Beranger; Sandrine Benghezal; Bérangère S Joly; Sophie Capdenat; Adeline Delton; Alain Stepanian; Paul Coppo; Agnès Veyradier Journal: Res Pract Thromb Haemost Date: 2020-12-15
Authors: Murtadha Al-Khabori; Faisal Alsayegh; Hasan Al Yaseen; Sabir Hussien; Amar Lal; Muna Al Rasheed; Mohammad Al Bader; Salam Al Kindi; Mahmoud Marashi Journal: Oman Med J Date: 2022-07-31