RHD VARIANTS INCLUDING REGULATORY TYPE RHNULL IN RUSSIANS
(Abstract release date: 05/19/16)
EHA Library. Golovkina L. 06/09/16; 135119; PB2219
Dr. Larisa Golovkina
Contributions
Contributions
Abstract
Abstract: PB2219
Type: Publication Only
Background
In clinical transfusions the D antigen of the Rhesus blood group system (BGS) is the third most relevant behind the A and B antigens of the ABO BGS Approximately 1%>2% of Caucasians show weak expression of D antigen on the RBC surface, which is caused by various types of mutations in the RHD gene. There are three D antigen variants in RH system: weak D, partial D and Del. It’s essential to know D variant distribution in each population in order to define whether a patient has a high or low probability to produce anti-D alloimmune antibodies during erythrocyte’s transfusion therapy.
Aims
to determine the distribution of D antigen variants in Russians.
Methods
Blood samples from 5100 Russians. 65 people have been investigated in our lab due to diminished D expression, discordant results with different anti-D reagents, or D negative samples with atypical Rh phenotype. D antigen was typed with 5 serological methods saline and indirect antiglobulin tests. The blood samples were serotyped for Rh C/c and Rh E/e antigens by corresponding Moabs. DNA extraction was followed by commercial SSP-PCR kits with primers for typing RHD, RHCE, 11 weak D types, partial D (BAG, Germany). Mutations in RHD (for confirmations of weak D type 3 - two cases and weak D type 15 - one case) and RHAG genes was detected by direct Taq cycle-sequencing, computer based sequence analysis and comparison with reference sequences from GenbankTM.
Results
In 2014-2015 years we revealed low expression of RhD antigen in 109 (2%) persons by screening. Genotyping 65 of them revealed six weak D types in 62 persons: weak D type 3 (47,7%; n=31; phenotypes – C+c+E-e+ in 28 cases, C+c-E-e+ in two and atypical one C-c+E-e+ ) and weak D type 1 (27,7%; n=18; phenotypes – C+c+E-e+) were the most frequent. Weak D type 2 was detected in 12,3% (n=8; phenotypes – C-c+E+e+), weak D type 15 – in 4,6% (n=3; phenotypes – C+c-E-e+, C+c+E-e+ and atypical one C-c+E+e+), weak D types 4.2 (DAR; phenotype C-c+E-e+) and weak D type 6 (phenotype C+c+E-e+) – each in one person (1,5% accordingly). All except 2 weak D types were identified by serological methods, and the most sensitive method was antiglobulin test in gel columns. Two cases with RhCe and RhcE phenotype were D-negative in all serological methods, and only molecular method allowed identifying weak D type 15. More one sample with diminished D expression was partial D variant DNB with phenotype C+c-E-e+. Two erythrocyte’s samples had true D-negative phenotype: one - RHDΨ with phenotype C-c+E-e+, and first in Russia (since 1926) we found regulatory type Rhnull with predicted phenotype D+C+c+E-e+ in woman with meningioma. RHAG sequencing shows polymorphism in exon 4 of the RHAG gene at position 571 (C > T) which leads to an earlier Stop-codon (amino acid position 191 [R191]) and the additional mutation at position 724 of exon 5 (G > A).
Conclusion
The distribution of D variants in Russians was identified. These data may help to improve the transfusion strategy of the D variant.
Session topic: E-poster
Keyword(s): Blood transfusion, Erythrocyte, Gene polymorphism
Type: Publication Only
Background
In clinical transfusions the D antigen of the Rhesus blood group system (BGS) is the third most relevant behind the A and B antigens of the ABO BGS Approximately 1%>2% of Caucasians show weak expression of D antigen on the RBC surface, which is caused by various types of mutations in the RHD gene. There are three D antigen variants in RH system: weak D, partial D and Del. It’s essential to know D variant distribution in each population in order to define whether a patient has a high or low probability to produce anti-D alloimmune antibodies during erythrocyte’s transfusion therapy.
Aims
to determine the distribution of D antigen variants in Russians.
Methods
Blood samples from 5100 Russians. 65 people have been investigated in our lab due to diminished D expression, discordant results with different anti-D reagents, or D negative samples with atypical Rh phenotype. D antigen was typed with 5 serological methods saline and indirect antiglobulin tests. The blood samples were serotyped for Rh C/c and Rh E/e antigens by corresponding Moabs. DNA extraction was followed by commercial SSP-PCR kits with primers for typing RHD, RHCE, 11 weak D types, partial D (BAG, Germany). Mutations in RHD (for confirmations of weak D type 3 - two cases and weak D type 15 - one case) and RHAG genes was detected by direct Taq cycle-sequencing, computer based sequence analysis and comparison with reference sequences from GenbankTM.
Results
In 2014-2015 years we revealed low expression of RhD antigen in 109 (2%) persons by screening. Genotyping 65 of them revealed six weak D types in 62 persons: weak D type 3 (47,7%; n=31; phenotypes – C+c+E-e+ in 28 cases, C+c-E-e+ in two and atypical one C-c+E-e+ ) and weak D type 1 (27,7%; n=18; phenotypes – C+c+E-e+) were the most frequent. Weak D type 2 was detected in 12,3% (n=8; phenotypes – C-c+E+e+), weak D type 15 – in 4,6% (n=3; phenotypes – C+c-E-e+, C+c+E-e+ and atypical one C-c+E+e+), weak D types 4.2 (DAR; phenotype C-c+E-e+) and weak D type 6 (phenotype C+c+E-e+) – each in one person (1,5% accordingly). All except 2 weak D types were identified by serological methods, and the most sensitive method was antiglobulin test in gel columns. Two cases with RhCe and RhcE phenotype were D-negative in all serological methods, and only molecular method allowed identifying weak D type 15. More one sample with diminished D expression was partial D variant DNB with phenotype C+c-E-e+. Two erythrocyte’s samples had true D-negative phenotype: one - RHDΨ with phenotype C-c+E-e+, and first in Russia (since 1926) we found regulatory type Rhnull with predicted phenotype D+C+c+E-e+ in woman with meningioma. RHAG sequencing shows polymorphism in exon 4 of the RHAG gene at position 571 (C > T) which leads to an earlier Stop-codon (amino acid position 191 [R191]) and the additional mutation at position 724 of exon 5 (G > A).
Conclusion
The distribution of D variants in Russians was identified. These data may help to improve the transfusion strategy of the D variant.
Session topic: E-poster
Keyword(s): Blood transfusion, Erythrocyte, Gene polymorphism
Abstract: PB2219
Type: Publication Only
Background
In clinical transfusions the D antigen of the Rhesus blood group system (BGS) is the third most relevant behind the A and B antigens of the ABO BGS Approximately 1%>2% of Caucasians show weak expression of D antigen on the RBC surface, which is caused by various types of mutations in the RHD gene. There are three D antigen variants in RH system: weak D, partial D and Del. It’s essential to know D variant distribution in each population in order to define whether a patient has a high or low probability to produce anti-D alloimmune antibodies during erythrocyte’s transfusion therapy.
Aims
to determine the distribution of D antigen variants in Russians.
Methods
Blood samples from 5100 Russians. 65 people have been investigated in our lab due to diminished D expression, discordant results with different anti-D reagents, or D negative samples with atypical Rh phenotype. D antigen was typed with 5 serological methods saline and indirect antiglobulin tests. The blood samples were serotyped for Rh C/c and Rh E/e antigens by corresponding Moabs. DNA extraction was followed by commercial SSP-PCR kits with primers for typing RHD, RHCE, 11 weak D types, partial D (BAG, Germany). Mutations in RHD (for confirmations of weak D type 3 - two cases and weak D type 15 - one case) and RHAG genes was detected by direct Taq cycle-sequencing, computer based sequence analysis and comparison with reference sequences from GenbankTM.
Results
In 2014-2015 years we revealed low expression of RhD antigen in 109 (2%) persons by screening. Genotyping 65 of them revealed six weak D types in 62 persons: weak D type 3 (47,7%; n=31; phenotypes – C+c+E-e+ in 28 cases, C+c-E-e+ in two and atypical one C-c+E-e+ ) and weak D type 1 (27,7%; n=18; phenotypes – C+c+E-e+) were the most frequent. Weak D type 2 was detected in 12,3% (n=8; phenotypes – C-c+E+e+), weak D type 15 – in 4,6% (n=3; phenotypes – C+c-E-e+, C+c+E-e+ and atypical one C-c+E+e+), weak D types 4.2 (DAR; phenotype C-c+E-e+) and weak D type 6 (phenotype C+c+E-e+) – each in one person (1,5% accordingly). All except 2 weak D types were identified by serological methods, and the most sensitive method was antiglobulin test in gel columns. Two cases with RhCe and RhcE phenotype were D-negative in all serological methods, and only molecular method allowed identifying weak D type 15. More one sample with diminished D expression was partial D variant DNB with phenotype C+c-E-e+. Two erythrocyte’s samples had true D-negative phenotype: one - RHDΨ with phenotype C-c+E-e+, and first in Russia (since 1926) we found regulatory type Rhnull with predicted phenotype D+C+c+E-e+ in woman with meningioma. RHAG sequencing shows polymorphism in exon 4 of the RHAG gene at position 571 (C > T) which leads to an earlier Stop-codon (amino acid position 191 [R191]) and the additional mutation at position 724 of exon 5 (G > A).
Conclusion
The distribution of D variants in Russians was identified. These data may help to improve the transfusion strategy of the D variant.
Session topic: E-poster
Keyword(s): Blood transfusion, Erythrocyte, Gene polymorphism
Type: Publication Only
Background
In clinical transfusions the D antigen of the Rhesus blood group system (BGS) is the third most relevant behind the A and B antigens of the ABO BGS Approximately 1%>2% of Caucasians show weak expression of D antigen on the RBC surface, which is caused by various types of mutations in the RHD gene. There are three D antigen variants in RH system: weak D, partial D and Del. It’s essential to know D variant distribution in each population in order to define whether a patient has a high or low probability to produce anti-D alloimmune antibodies during erythrocyte’s transfusion therapy.
Aims
to determine the distribution of D antigen variants in Russians.
Methods
Blood samples from 5100 Russians. 65 people have been investigated in our lab due to diminished D expression, discordant results with different anti-D reagents, or D negative samples with atypical Rh phenotype. D antigen was typed with 5 serological methods saline and indirect antiglobulin tests. The blood samples were serotyped for Rh C/c and Rh E/e antigens by corresponding Moabs. DNA extraction was followed by commercial SSP-PCR kits with primers for typing RHD, RHCE, 11 weak D types, partial D (BAG, Germany). Mutations in RHD (for confirmations of weak D type 3 - two cases and weak D type 15 - one case) and RHAG genes was detected by direct Taq cycle-sequencing, computer based sequence analysis and comparison with reference sequences from GenbankTM.
Results
In 2014-2015 years we revealed low expression of RhD antigen in 109 (2%) persons by screening. Genotyping 65 of them revealed six weak D types in 62 persons: weak D type 3 (47,7%; n=31; phenotypes – C+c+E-e+ in 28 cases, C+c-E-e+ in two and atypical one C-c+E-e+ ) and weak D type 1 (27,7%; n=18; phenotypes – C+c+E-e+) were the most frequent. Weak D type 2 was detected in 12,3% (n=8; phenotypes – C-c+E+e+), weak D type 15 – in 4,6% (n=3; phenotypes – C+c-E-e+, C+c+E-e+ and atypical one C-c+E+e+), weak D types 4.2 (DAR; phenotype C-c+E-e+) and weak D type 6 (phenotype C+c+E-e+) – each in one person (1,5% accordingly). All except 2 weak D types were identified by serological methods, and the most sensitive method was antiglobulin test in gel columns. Two cases with RhCe and RhcE phenotype were D-negative in all serological methods, and only molecular method allowed identifying weak D type 15. More one sample with diminished D expression was partial D variant DNB with phenotype C+c-E-e+. Two erythrocyte’s samples had true D-negative phenotype: one - RHDΨ with phenotype C-c+E-e+, and first in Russia (since 1926) we found regulatory type Rhnull with predicted phenotype D+C+c+E-e+ in woman with meningioma. RHAG sequencing shows polymorphism in exon 4 of the RHAG gene at position 571 (C > T) which leads to an earlier Stop-codon (amino acid position 191 [R191]) and the additional mutation at position 724 of exon 5 (G > A).
Conclusion
The distribution of D variants in Russians was identified. These data may help to improve the transfusion strategy of the D variant.
Session topic: E-poster
Keyword(s): Blood transfusion, Erythrocyte, Gene polymorphism
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