BACKGROUND AND OBJECTIVES: One to two per cent of patients in need of red cell transfusion carry irregular antibodies to red blood cell (RBC) antigens and have to be supplied with specially selected blood units. To be able to respond to those requests, blood centres have to screen a significant number of donors for a variety of antigens serologically, which is a costly and through the shortage of reagents, also limited procedure. To make this procedure more efficient, the Austrian Red Cross has developed a genotyping assay as an alternative approach for high throughput RBC typing. MATERIALS AND METHODS: A multiplex polymerase chain reaction (PCR) assay was designed for typing 35 RBC antigens in six reaction mixes. The assay includes both common as well as high-frequency-alleles: MNS1, MNS2, MNS3 and MNS4; LU1, LU2, LU8 and LU14; KEL1, KEL2, KEL3, KEL4, KEL6, KEL7, KEL11, KEL17 and KEL21; FY1, FY2, FYB(WK) and FY0 (FYB(ES)); JK1 and JK2; DI1, DI2, DI3 and DI4; YT1 and YT2; DO1 and DO2; CO1 and CO2; IN1 and IN2. The assay was validated using 370 selected serologically typed samples. Subsequently 6000 individuals were screened to identify high frequency antigen (HFA)-negative donors and to facilitate the search for compatible blood for alloimmunized patients. RESULTS: All controls showed complete concordance for the tested markers. The screening of 6000 donors revealed 57 new HFA-negative donors and the blood group database was extended by approximately 210,000 results. CONCLUSION: The study shows that in practice, this high-throughput genotyping assay is feasible, fast and provides reliable results. Compared to serological testing, this molecular approach is also very cost-efficient.
BACKGROUND AND OBJECTIVES: One to two per cent of patients in need of red cell transfusion carry irregular antibodies to red blood cell (RBC) antigens and have to be supplied with specially selected blood units. To be able to respond to those requests, blood centres have to screen a significant number of donors for a variety of antigens serologically, which is a costly and through the shortage of reagents, also limited procedure. To make this procedure more efficient, the Austrian Red Cross has developed a genotyping assay as an alternative approach for high throughput RBC typing. MATERIALS AND METHODS: A multiplex polymerase chain reaction (PCR) assay was designed for typing 35 RBC antigens in six reaction mixes. The assay includes both common as well as high-frequency-alleles: MNS1, MNS2, MNS3 and MNS4; LU1, LU2, LU8 and LU14; KEL1, KEL2, KEL3, KEL4, KEL6, KEL7, KEL11, KEL17 and KEL21; FY1, FY2, FYB(WK) and FY0 (FYB(ES)); JK1 and JK2; DI1, DI2, DI3 and DI4; YT1 and YT2; DO1 and DO2; CO1 and CO2; IN1 and IN2. The assay was validated using 370 selected serologically typed samples. Subsequently 6000 individuals were screened to identify high frequency antigen (HFA)-negative donors and to facilitate the search for compatible blood for alloimmunized patients. RESULTS: All controls showed complete concordance for the tested markers. The screening of 6000 donors revealed 57 new HFA-negative donors and the blood group database was extended by approximately 210,000 results. CONCLUSION: The study shows that in practice, this high-throughput genotyping assay is feasible, fast and provides reliable results. Compared to serological testing, this molecular approach is also very cost-efficient.
Authors: Willy A Flegel; Shirley L De Castilho; Margaret A Keller; Ellen B Klapper; Joann M Moulds; France Noizat-Pirenne; Nadine Shehata; Gary Stack; Maryse St-Louis; Christopher A Tormey; Dan A Waxman; Christof Weinstock; Silvano Wendel; Gregory A Denomme Journal: Blood Transfus Date: 2015-12-21 Impact factor: 3.443
Authors: Genghis H Lopez; Rhiannon S Mcbean; Brett Wilson; Darryl L Irwin; Yew-Wah Liew; Catherine A Hyland; Robert L Flower Journal: Blood Transfus Date: 2014-06-05 Impact factor: 3.443
Authors: Christoph Gassner; Frauke Degenhardt; Stefan Meyer; Caren Vollmert; Nadine Trost; Kathrin Neuenschwander; Yvonne Merki; Claudia Portmann; Sonja Sigurdardottir; Antigoni Zorbas; Charlotte Engström; Jochen Gottschalk; Soraya Amar El Dusouqui; Sophie Waldvogel-Abramovski; Emmanuel Rigal; Jean-Daniel Tissot; Caroline Tinguely; Simon M Mauvais; Amira Sarraj; Daniel Bessero; Michele Stalder; Laura Infanti; Andreas Buser; Jörg Sigle; Tina Weingand; Damiano Castelli; Monica C Braisch; Jutta Thierbach; Sonja Heer; Thomas Schulzki; Michael Krawczak; Andre Franke; Beat M Frey Journal: Transfus Med Hemother Date: 2018-07-10 Impact factor: 3.747