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ABSTRACT Introduction: Data on the prevalence of clinically significant antigens (Rh, Kell, Kidd, Duffy, MNSs, Lewis, P and Lutheran) among the Indian donor population is sparse. Objective: This prospective study was aimed at determining the prevalence of 21 clinically significant antigens for the first time in the South Indian donor population. Method: A total of 672 regular O group blood donors were enrolled for Rh (C, c, E, e) and Kell (K) antigens typing. Of these, 188 donors were typed for other clinically significant antigens (k, Kpa, Kpb, Jka, Jkb, Fya, Fyb, M, N, S, s, P1, Lea, Leb, Lua and Lub). Results: Antigen frequencies were expressed in percentages. In our study, R1R1 and rr were the most common phenotypes among D+ and D− donors, respectively. Among the Rh antigens, the e antigen was expressed by 97.5% and 100% of D+ and D− donors, respectively. The K antigen was found in only 0.15% of donors. In the Duffy and Kidd blood group system, Fy (a+b+) and Jk (a+b+) were the most frequent phenotypes, respectively. In the MNSs blood group system, M+N+ and S−s+ were the most common phenotypes reported. The Le (a−b+) was found to be the phenotype with the highest prevalence in the Lewis blood group system. The Lu (a−b+) was the only phenotype found in the Lutheran blood group system. Conclusion: Knowledge regarding the prevalence of antigens in a given population is essential in developing cost-effective in-house panels and a rare donor registry comprising donors typed negative for a high-frequency antigen or a combination of common multiple antigens. Introduction Rh, (Rh sparse Objective 2 Method 67 C, C (C c E (K typing these 18 k, k (k Kpa Kpb Jka Jkb Fya Fyb M N S s P1 Lea Leb Lub. Lub . Lub) Results percentages RR R R1R D respectively 975 97 5 97.5 100 015 0 15 0.15 system a+b+ ab b (a+b+ MN M+N Ss S−s reported a−b+ (a−b+ Conclusion costeffective cost effective inhouse house highfrequency high frequency 6 1 9 97. 10 01 0.1 a+b (a+b a−b (a−b 0.