Comparison of the four methods regularly used for pre-transfusion antibody screening Chun-Chang Lee 1 , Lei-Fa Chang 2 , Hai-Lung Wang 2 1 Cheng Hsin General Hospital Department of Clinical Pathology-Blood Bank, 2 Yuan-Pei Technical University Conclusion ： By the comparison, we demonstrated the advantages of By the comparison, we demonstrated the advantages of Sanquin Cellbind were high sensitivity and low false positive . It could replace the time-consuming PEG and less sensitive LISS to be a regular examination. Moreover, it could be used for double check when the result by MP test was not clear and definite. At present, Sanquin Cellbind is a standardized and automated system. Using the system, time and labors can be saved and systemic error can be avoided.
Comparison of the four methods regularly used for pre-transfusion antibody screening Background ： The goal of this study is to compare four methods commonly used in serology, including manual polybrene (MP), Polyethylene glycol antiglobulin (PEG), Low ionic strength saline (LISS) and Sanquin Cellbind immunofixation column, which were used to screen antibodies in clinical tests. We tested the sensitivity of different were used to screen antibodies in clinical tests. We tested the sensitivity of different antigen-antibodies respectively and compare the specificity of four methods. Then, we also compared the reactions of four methods with known positive specimens and compare their specificity with unknown specimens.
Comparison of the four methods regularly used for pre-transfusion antibody screening Material ： Sanquin rare blood grouping reagents (human based), Sanquin screening panel 123 cell, 100 specimens known with antibodies and 187 unknown specimens. Method ： 1. To compare the sensitivity and specificity of four methods by Sanquin rare blood grouping reagents and Sanquin sreening panel 123 cell. 2. To test the known positive specimens with Sanquin screening panel 123 and compare the specificity of different methods. 3. To identify the antibodies of unknown specimens by Sanquin different methods. 3. To identify the antibodies of unknown specimens by Sanquin screening panel 123 (the blind test).
Comparison of the four methods regularly used for pre-transfusion antibody screening Results ： From Table1, Sanquin Cellbind had identified the antibodies, especially anti-D, anti-C, anti-E, anti-e and anti-c. All of them had a titer of at least 1:1024. There were 8 among the 13 antibodies had the highest titer. According to Table2, the specificity of MP was the highest and the range was 60%~100%. But the range of the other methods were not stable because the amounts of specimens were not enough. After identifying the 100 positive specimens by four methods, we found the positive rate were 91%, 66%, 81% and positive specimens by four methods, we found the positive rate were 91%, 66%, 81% and 79%, respectively (Table3). The highest was MP, followed by PEG and Sanquin Cellbind and the lowest was LISS. From Table4, there were 6 positive reactions by MP and Sanquin Cellbind and 4 positive reactions by LISS and PEG. This result showed the sensitivity of MP and Sanquin Cellbind was higher in clinical tests.
Comparison of the four methods regularly used for pre-transfusion antibody screening Table1 the comparison of the sensitivity between four methods The type MP LISS PEG Cellbind of antibody anti-Jka 1:128 1:32 1:128 *1:512 anti-Jkb 1:128 1:16 1:128 *1:128 anti-K 1:128 1:64 1:512 1:64 anti-k 1:16 1:32 1:128 1:64 anti-Fya 1:8 1:16 1:16 *1:32 anti-Fyb 1:16 1:32 1:64 1:32 anti-S 1:128 1:512 1:512 1:128 anti-s 1:32 1:64 1:128 1:32 anti-D 1:4096 1:1024 1:1024 *1:4096 anti-C 1:512 1:256 1:512 *1:2048 anti-E 1:2048 1:512 1:2048 *1:2048 anti-e 1:1024 1:256 1:1024 *1:1024 anti-c 1:2048 1:512 1:512 *1:2048
Comparison of the four methods regularly used for pre-transfusion antibody screening Table2 the comparison of antibody detection between four methods Antibodies N MP LISS PEG Cellbind P1 2 2 (100%) 1 (50%) 1 (50%) 1 (50%) E 13 13 (100%) 7 (54%) 12 (92%) 13 (100%) M 21 21 (100%) 16 (76%) 20 (95%) 15 (71%) Leb 2 2 (100%) 2 (100%) 2 (100%) 0 (0%) Lea 6 6 (100%) 5 (83%) 6 (100%) 6 (100%) Fyb 2 2 (100%) 2 (100%) 1 (50%) 2 (100%) c 1 1 (100%) 0 (0%) 0 (0%) 1 (100%) Jkb 2 2 (100%) 2 (100%) 2 (100%) 2 (100%) N 2 2 (100%) 1 (50%) 2 (100%) 2 (100%) Jka 4 3 (75%) 3 (75%) 3 (75%) 3 (75%) S 4 3 (75%) 3 (75%) 3 (75%) 3 (75%) E+c 15 9 (60%) 4 (27%) 7 (47%) 8 (53%) C+e 3 3 (100%) 0 (0%) 0 (0%) 0 (0%) Jkb+E 1 1 (100%) 0 (0%) 1 (100%) 1 (100%) Lea+leb 1 1 (100%) 1 (100%) 1 (100%) 1 (100%) D 4 4 (100%) 4 (100%) 4 (100%) 4 (100%) Mia 15 14 (93%) 13 (87%) 14 (93%) 15 (100%) Dia 2 2 (100%) 2 (100%) 2 (100%) 2 (100%) 100
Comparison of the four methods regularly used for pre-transfusion antibody screening Table3 The comparison of incidence of known positive specimens N MP LISS PEG Cellbind 100 91 (91%) 66 (66%) 81 (81%) 79 (79%) Table4 The comparison of positive incidence of unknown specimens N MP LISS PEG Cellbind 187 6 (3.2%) 4 (2.1%) 4 (2.1%) 6 (3.2%)
Comparison of the four methods regularly used for pre-transfusion antibody screening Discussion ： According to the results tested by commercial serum reagent, Sanquin Cellbind had high sensitivity and could be used in regular examination . The result of antibody detection showed four methods had different specificity to each antibody. Because MP was the regular method in Taiwan, the known specimens have been tested once in the hospital. MP showed the better reproducibility. The other method had its advantage to some antibodies, such as PEG to antibody M and Cellbind to antibody E, Lea or Mia. But except MP method, the error range of specificity were too large in the tests with positive specimens. We considered the different amounts of the positive specimens was the major reason. Some specimens were so few that the error range specimens was the major reason. Some specimens were so few that the error range became too big. If we could collect more than 10 specimens, the statistics would be more representative. On the other hand, there were still some false negative reactions in this test. It might be the reason that the titer of antibodies in these specimens became weaker by the time. Some reactions have been +1 or trace reactions in the hospital but disappeared in this study. Additionally, the reactions of some antibodies were weak from the beginning. After frozen storage for a long time, the antibodies were destroyed and could not be detected again. In the blind test, a. It indicated Sanquin Cellbind was able to be used regularly in clinical test as MP. Even it could replace LISS or PEG to double check the results of MP.
Comparison of the four methods regularly used for pre-transfusion antibody screening Conclusion ： By the comparison, we demonstrated the advantages of Sanquin Cellbind were high sensitivity and low false positive . It could replace the time- consuming PEG and less sensitive LISS to be a regular examination. Moreover, it could be used for double check when the result by MP test was not clear and definite. At present, Sanquin Cellbind is a standardized and automated system. definite. At present, Sanquin Cellbind is a standardized and automated system. Using the system, time and labors can be saved and systemic error can be avoided.
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