ABSTRACT
【Objective】 To identify one blood sample with ambiguous antibody screening result, explore the identification strategy of complex antibodies and provide blood transfusion plan. 【Methods】 Blood typing and antibody screening were performed by serological test. The genotyping of Diego blood group was carried out by gene detection. Absorption and elution test were designed to identify the complex antibodies. 【Results】 Serological test showed that the patient′s blood type was B, ccDEE, Jk(a-b+ ), Le(a-b+ ), Fy(a+ b-), Di(a+ b-). The results of genotyping showed that the Diego genotype of the patient was Di (a+ b-). Based on the above results, absorption and elution tests were designed and IgG anti-Dib, IgG anti-Ce, and IgG anti-Jka antibodies were detected in serum of the patient. Combined with the use of immunoglobulin and targeted blood transfusion measures, successful treatment was provided for the patient. 【Conclusion】 For the identification of antibodies against high frequency antigen combined with multiple antibodies, a variety of experimental techniques and methods should be combined, and the experimental scheme should be designed flexibly.
ABSTRACT
BACKGROUND: Wra is a low-incidence antigen, which is antithetical to the high prevalence red blood cell antigen, Wrb. Anti-Wra is a naturally occurring antibody that is found in approximately 1-2% of blood donors. The aim of this study was to determine the frequency of Wra and anti-Wra in Brazilian blood donors.METHODS: A total of 1662 Brazilian blood donors were molecularly analyzed using the SNaPshot methodology to determine the WR*A/B alleles and to predict the frequency of the Wra antigen. To detect the anti-Wra, samples from 1049 blood donors were analyzed using a gel test with Wr(a+) red blood cells. The serum was treated with dithiothreitol (DTT) to determine the immunoglobulin classes. Immunoglobulin (Ig)-G isotype classification was performed in a gel test using the IgG1/IgG3 card. A monocyte monolayer assay was employed to predict the clinical significance of IgG anti-Wra.RESULTS: Of the 1662 donors, only one sample had the DI*02.03 allele in heterozygous predicting the Wr(a+b+) phenotype. Anti-Wra was detected in 34 (3.24%) samples, 64.7% in females and 35.3% in males. Regarding the immunoglobulin class, eight (23.5%) cases of anti-Wra were classified as IgG and 26 (76.5%) as IgM. Of the eight cases of IgG anti-Wra, four were IgG1, two were IgG3 and three anti-Wra were not IgG3 or IgG1, and thus probably IgG2 or IgG4. The results of the monocyte monolayer assay showed that IgG anti-Wra might be of clinical significance.CONCLUSION: This study shows a very low frequency (0.06%) of the Wra antigen in Brazilian blood donors. Additionally, it shows that the frequency of anti-Wra in this population is higher than previously reported.
Subject(s)
Humans , Blood Donors , Blood Group Antigens , Gene FrequencyABSTRACT
BACKGROUND: The Diego blood group system consists of two independent pairs of antigens, Dia and Dib. Immunization to Dia or Dib is clinically significant, because anti-Dia and anti-Dib may cause hemolytic transfusion reactions of transfused incompatible red cells or hemolytic disease of the newborn. At the nucleotide level, the difference between the Di a and Di b alleles is a single-base change of exon 19 that results in the substitution of leucine (CTG) for proline (CCG) at position 854. METHODS: Peripheral blood was collected from 116 patients. DNA was isolated from 50 L of blood. PCR was performed with previously described primers by Bruce et al (S22: 5'-GTC ACGTCGCTCAGCGG, AS13: 5'-GACCTTCCTCCTCATCAA). The 5 L of PCR products were digested by Nae I. We analyzed 10ul of each digested PCR product by electrophoresis on 1.5 % agarose gel with ethidium bromide staining. RESULTS: A concordance rate of 100 percent was observed between genotyping and phenotyping (105 Di (a-b+), 11 Di (a+b+)). CONCLUSIONS: This method can be effectively used for the Diego typing and is particularly useful in cases where the serological typing method is difficult as in autoimmune hemolytic anemia.