1.Genotyping for Kidd, Kell, Duffy, Scianna, and RHCE blood group antigens polymorphisms in Jiangsu Chinese Han.
Zhong LIU ; Rong ZENG ; Qing CHEN ; Min LI ; Guang-yao SHI ; Peng WEI ; Cheng-yin HUANG ; Rong-cai TANG ; Jun SUN ; Xue-guang ZHANG
Chinese Medical Journal 2012;125(6):1076-1081
<p>BACKGROUNDMolecular testing is more precise compared to serology and has been widely used in genotyping blood group antigens. Single nucleotide polymorphisms (SNPs) of blood group antigens can be determined by the polymerase chain reaction with sequence specific priming (PCR-SSP) assay. Commercial high-throughput platforms can be expensive and are not approved in China. The genotype frequencies of Kidd, Kell, Duffy, Scianna, and RhCE blood group antigens in Jiangsu province were unknown. The aim of this study is sought to detect the genotype frequencies of Kidd, Kell, Duffy, Scianna, and RhCE antigens in Jiangsu Chinese Han using molecular methods with laboratory developed tests.p><p>METHODSDNA was extracted from EDTA-anticoagulated blood samples of 146 voluntary blood donors collected randomly within one month. Standard serologic assay for red blood cell antigens were also performed except the Scianna blood group antigens. PCR-SSP was designed to work under one PCR program to identify the following SNPs: JK1/JK2, KEL1/KEL2, FYA/FYB, SC1/SC2, C/c and E/e.p><p>RESULTSSerologic antigen results were identical to the phenotypes that were predicted from genotyping results. The allele frequencies for Jk*01 and Jk*02 were 0.51 and 0.49, respectively; for Fy*A and Fy*B 0.94 and 0.06; for RHCE*C and RHCE*c 0.68 and 0.32; and for RHCE*E and RHCE*e 0.28 and 0.72. Among 146 blood donors, all were KEL*02/KEL*02 and SC*01/SC*01, indicating allele frequencies for KEL*02 and SC*01 close to 1.00.p><p>CONCLUSIONSThe use of PCR-SSP working under the same condition for testing multiple antigens at the same time is practical. This approach can be effective and cost-efficient for small-scale laboratories and in developing counties. These molecular tests can be also used for identifying rare blood types.p>
Blood Group Antigens
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genetics
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Butyrophilins
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China
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ethnology
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Duffy Blood-Group System
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genetics
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Gene Frequency
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Genotype
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Humans
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Kell Blood-Group System
;
genetics
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Kidd Blood-Group System
;
genetics
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Polymerase Chain Reaction
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Polymorphism, Single Nucleotide
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Rh-Hr Blood-Group System
;
genetics
3.Analysis of Patients' Irregular Antibody Screening and Identification Results before Blood Transfusion.
Hui LI ; Huan-Ming XU ; Yi ZHANG ; Ji-Xiang CUI
Journal of Experimental Hematology 2015;23(3):861-865
<p>OBJECTIVETo analyze the results of irregular antibody screening and identification among patients before blood transfusion, and to investigate the specific distribution of irregular antibodies and the distribution regularity in different diseases.p><p>METHODSChoosing the patients intended to be transfused in our hospital from January 1, 2009 to December 31, 2013 years, micro-column gel technique was used to screen the irregular antibodies of those receptors and to identify the antibody specificity of the positive specimens.p><p>RESULTSAmong 44194 patients, 137 patients were with irregular antibody positive and their positive rate was 0.31%, among them 33 cases were male and accounted for 0.18% in the studied males; the 104 cases were females and accounted for 0.40% in all the studied females. The difference of sex distribution was statistically significant (X2=15.38, P<0.05). In the irregular antibody screening positive patients, patients with transfusion or pregnancy history were 129 cases, and the patients without transfusion or pregnancy history were 8 cases. In the irregular antibody screening positive patients, the main antibody of 54 cases belongs to Rh blood type system, accounting for 39.42%; The main antibody of 37 cases belongs to MNS blood type system, accounting for 27.01%; while the 30 cases belong to Lewis blood type system, accounting for 21.90%. According to the classification of diseases, the irregular antibody screening-positive patients with tumors were ranked in the highest rate at 5.96‰, the secondary hemorrhage of digestive tract and chronic renal failure were ranked at the rate of 3.28‰ and 3.19‰. The difference of positive rates between diseases was statistically significant (χ2=19.33, P<0.05).p><p>CONCLUSIONIrregular antibody screening before blood transfusion is necessary, which can discover the irregular antibodies of clinical significance, especially for patients with tumors and the other patients with the history of frequent blood transfusions or multiple pregnancies. Antibody screening is a useful warning signal, as it ensures the safety of blood transfusions.p>
Antibodies
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Blood Group Antigens
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Blood Grouping and Crossmatching
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Blood Transfusion
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Female
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Gastrointestinal Tract
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Humans
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Lewis Blood-Group System
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Male
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Pregnancy
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Rh-Hr Blood-Group System
4.Application of multiplex PCR for the screening of genotyping system for the rare blood groups Fy(a-), s-,k-,Di(b-) and Js(b-).
Wei JIAO ; Li XIE ; Hailan LI ; Jiao LAN ; Zhuning MO ; Ziji YANG ; Fei LIU ; Ruiping XIAO ; Yunlei HE ; Luyi YE ; Ziyan ZHU
Chinese Journal of Medical Genetics 2014;31(2):242-246
<p>OBJECTIVETo screen rare blood groups Fy(a-), s-, k-, Di(b-) and Js(b-) in an ethnic Zhuang population.p><p>METHODSSequence-specific primers were designed based on single nucleotide polymorphism (SNP) sites of blood group antigens Fy(b) and s. A specific multiplex PCR system I was established. Multiplex PCR system II was applied to detect alleles antigens Di(b), k, Js(b)1910 and Js(b) 2019 at the same time. The two systems was were used to screen for rare blood group antigens in 4490 randomly selected healthy donors of Guangxi Zhuang ethnic origin.p><p>RESULTSWe successfully made the multiplex PCR system I. We detected the rare blood group antigens using the two PCR system. There are five Fy(a-), three s(-), two Di(b-) in 4490 Guangxi zhuang random samples. The multiplex PCR system I has achieved good accuracy and stability. With multiplex PCR systems I and II, 4490 samples were screened. Five Fy(a-), three s(-) and two Di(b-) samples were discovered.p><p>CONCLUSIONMultiplex PCR is an effective methods, which can be used for high throughput screening of rare blood groups. The rare blood types of Guangxi Zhuang ethnic origin obtained through the screening can provide valuable information for compatible blood transfusion. Through screening we obtained precious rare blood type materials which can be used to improve the capability of compatible infusion and reduce the transfusion reactions.p>
Blood Group Antigens
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genetics
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Duffy Blood-Group System
;
genetics
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Genotype
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Humans
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Multiplex Polymerase Chain Reaction
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methods
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Receptors, Cell Surface
;
genetics
5.Blood group phenotype frequencies in blood donors from a tertiary care hospital in north India.
Nitin AGARWAL ; Rakesh Mohan THAPLIYAL ; Kabita CHATTERJEE
Blood Research 2013;48(1):51-54
BACKGROUND: Knowledge about the frequency of red blood cell-antigen phenotypes in a population can be helpful in the creation of a donor data bank for the preparation of indigenous cell panels and for providing antigen-negative compatible blood to patients with multiple alloantibodies. METHODS: ABO and RhD blood grouping was performed on 9,280 continuous voluntary and replacement donors. For other rare blood groups, 508 ACD blood samples were obtained from the donors at the Blood Bank of the Department of Transfusion Medicine, All India Institute of Medical Sciences (AIIMS), New Delhi, India. Blood group antigens were determined by tube method using anti-sera (Bio-Rad, USA), and the phenotype frequencies were expressed as percentages. RESULTS: Group B (37.39%) was the most common, followed by group O (31.85%). R1R1 and rr were the most common phenotypes amongst Rh positive and Rh negative groups, respectively. A rare phenotype R2Rz was found in one donor. For Kidd and Duffy blood group systems, Jk (a+b+) and Fy (a+b+) were the most common phenotypes (46.06% and 48.03%, respectively). The most common phenotypes for MNSs, Lu, and Kell blood groups were M+N+, S-s+, Lu (a-b+), and K-k+, respectively. A very rare case of Fy (a-b-) and Jk (a-b-) was found in a single donor. CONCLUSION: This study is the first small step to create a rare donor data bank and to prepare indigenous cell panels to provide compatible blood to all multi-transfused alloimmunized patients.
Blood Banks
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Blood Donors
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Blood Group Antigens
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Blood Grouping and Crossmatching
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Duffy Blood-Group System
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Humans
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India
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Phenotype
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Tertiary Healthcare
;
Tissue Donors
6.Blood group phenotype frequencies in blood donors from a tertiary care hospital in north India.
Nitin AGARWAL ; Rakesh Mohan THAPLIYAL ; Kabita CHATTERJEE
Blood Research 2013;48(1):51-54
BACKGROUND: Knowledge about the frequency of red blood cell-antigen phenotypes in a population can be helpful in the creation of a donor data bank for the preparation of indigenous cell panels and for providing antigen-negative compatible blood to patients with multiple alloantibodies. METHODS: ABO and RhD blood grouping was performed on 9,280 continuous voluntary and replacement donors. For other rare blood groups, 508 ACD blood samples were obtained from the donors at the Blood Bank of the Department of Transfusion Medicine, All India Institute of Medical Sciences (AIIMS), New Delhi, India. Blood group antigens were determined by tube method using anti-sera (Bio-Rad, USA), and the phenotype frequencies were expressed as percentages. RESULTS: Group B (37.39%) was the most common, followed by group O (31.85%). R1R1 and rr were the most common phenotypes amongst Rh positive and Rh negative groups, respectively. A rare phenotype R2Rz was found in one donor. For Kidd and Duffy blood group systems, Jk (a+b+) and Fy (a+b+) were the most common phenotypes (46.06% and 48.03%, respectively). The most common phenotypes for MNSs, Lu, and Kell blood groups were M+N+, S-s+, Lu (a-b+), and K-k+, respectively. A very rare case of Fy (a-b-) and Jk (a-b-) was found in a single donor. CONCLUSION: This study is the first small step to create a rare donor data bank and to prepare indigenous cell panels to provide compatible blood to all multi-transfused alloimmunized patients.
Blood Banks
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Blood Donors
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Blood Group Antigens
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Blood Grouping and Crossmatching
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Duffy Blood-Group System
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Humans
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India
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Phenotype
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Tertiary Healthcare
;
Tissue Donors
7.Effect of 2Me, DTT, ZAAP and Enzyme on JMH Antigen on the Surface of Human Erythrocytes.
Zhao-Hu YUAN ; Hui-Kuan YANG ; Xiao-Jie CHEN ; Ya-Ming WEI
Journal of Experimental Hematology 2019;27(3):920-924
OBJECTIVE:
To explore the the effects of 2-Me, DTT, papain, pineapple protease and ZZAP on the antigenicity of JMH antigen of human red blood cells (RBC) surface.
METHODS:
Firstly, human RBC were treated with 2-Me, DTT, pineapple protease, papain and ZZAP reagents, respectively. The antigenicity of JMH antigen on human RBC surface was detected and analyzed by flow cytometry.
RESULTS:
Flow cytometric analysis found that compared with level before treatment, the antigenicity of JMH antigen on RBC surface was significantly reduced after 2-Me treatment, the positive rate of JMH antigen: 69.5%±4.5% vs 56.5%±3.4% (t=12.44, P<0.01); fluorescence intensity: 4906±317 vs 3003±165 (t=11.84, P<0.01). The antigenicity of JMH antigen on RBC surface significantly increased after DTT treatment, showing the positive rate of JMH antigen: 61.7%±3.8% vs 75.5±4.9% (t=16.57, P<0.01), fluorescence intensity: 4044±294 vs 4854±319 (t=15.46, P<0.01). However, both bromelain and papain could significantly reduce the antigenicity of JMH antigen on the RBC surface, Bromelain: the positive rate of JMH antigen: 62.2%±3.8% vs 8.8%±1.2% (t=26.44, P<0.01), fluorescence intensity: 4263±273 vs 1444±212 (t=19.27, P<0.01); Papain: the positive rate of JMH antigen: 62.8%±3.6% vs 8.8%±1.5% (t=21.38, P<0.01), fluorescence intensity: 4389±284 vs 1458±230 (t=17.49, P<0.01). The flow cytometric analysis revealed that ZZAP treatment significantly reduced the antigenicity of JMH antigen on the RBC surface, the positive rate of JMH antigen: 62.2%±4.4% vs 48.2%±4.1% (t=14.87, P<0.01), fluorescence intensity: 4106±263 vs 2063±175 (t=17.49, P<0.01).
CONCLUSION
The treatment with 2-Me can reduce the antigenicity of JMH antigen on human RBC surface. The antigenicity of JMH antigen on human RBC surface increased after DTT treatment. The antigenicity of JMH antigen on human RBC surface significantly reduces after the treatment with pineapple protease or papain. ZZAP treatment can reduce the antigenicity of JMH antigen on the RBC surface.
Blood Group Antigens
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Erythrocytes
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Flow Cytometry
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Humans
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Rh-Hr Blood-Group System
8.Analysis of Irregular Blood Group Antibody Distribution and Blood Transfusion Efficacy in Patients with Malignant Tumor.
Dong REN ; Hong-Bin ZHAO ; Xiao-Jun GUO ; Xue-Hua HE
Journal of Experimental Hematology 2023;31(1):209-214
OBJECTIVE:
To investigate the distribution of irregular blood group antibodies in patients with malignant tumors, and to analyze the relationship between it and efficacy of blood transfusion in patients.
METHODS:
5 600 patients with malignant tumors treated in Shanxi Bethune Hospital from January 2019 to December 2021 were selected as the research subjects. All patients received blood transfusion, and cross matching test was conducted before blood transfusion, irregular antibody results of patients were tested; the irregular distribution of blood group antibodies was observed, and the relationship between it and efficacy of blood transfusion in patients was analyzed.
RESULTS:
Among 5 600 patients with malignant tumors, 96 cases were positive for irregular antibody, and the positive rate was 1.71%; the main blood group systems involved in the irregular antibody positive of 96 patients with malignant tumors were RH, MNSs and Duffy system, among which Rh blood group was the most common, and the proportion of anti-E was the highest; among the malignant tumor patients with positive blood group irregular antibody, the proportion of female was higher than that of male; the proportion of patients aged >60 years was the highest, followed by patients aged >40 and ≤50 years, and the proportion of patients aged 18-30 years was the lowest; the patients with positive blood group irregular antibody were mainly in blood system (including lymphoma), digestive system, reproductive and urinary system; the positive rate of irregular antibody of patients in the ineffective group was higher than that of patients in the effective group, the difference was statistically significant (<i>Pi><0.05). Logistic regression analysis results showed that, irregular antibody positive was a risk factor for ineffective blood transfusion in patients with malignant tumor (OR>1, <i>Pi><0.05).
CONCLUSION
The irregular blood group antibody positive of patients with malignant tumor are mostly female, and the proportion of patients aged >60 is the highest, which is mainly distributed in malignant tumors of blood system, digestive system and urogenital system, and the positive blood group irregular antibody is related to the efficacy of blood transfusion in patients.
Humans
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Male
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Female
;
Blood Transfusion
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Blood Group Antigens
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Rh-Hr Blood-Group System
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Antibodies
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Neoplasms/therapy*
;
Isoantibodies
9.Detecting ABO blood type of bloodstain with fluorescent antibody method.
Rui-Heng WANG ; Li-Qiang YAN ; Wei-Jian YU ; Xiao-Wei LIU ; Shu-xian QU
Journal of Forensic Medicine 2007;23(2):127-129
UNLABELLED:
OBJECTIVE To explore the advantage and feasibility of fluorescent antibody method for detection of blood type in biological material.
METHODS:
According to theory of specific binding of antigen and antibody, at first the anti-A monoclonal antibody (MA) and anti-B MA were labeled with the fluorescent, then fluorescent-labeled antibodies (FLA) were bound with corresponding biological material (such as bloodstain) in the optimum condition, finally the ABO blood type of bloodstain was determined under microscope fluorescent.
RESULTS:
The fluorescent antibody method is highly sensitive, accurate and simple.
CONCLUSION
The fluorescent antibody method is an accurate and reliable method for detection of ABO blood type in biological material.
ABO Blood-Group System/immunology*
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Antibodies, Monoclonal/blood*
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Antigen-Antibody Reactions
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Blood Group Antigens/blood*
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Blood Stains
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Fluorescent Antibody Technique/methods*
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Forensic Medicine/methods*
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Humans
;
Sensitivity and Specificity
10.Molecular Mechanism of a Rhesus D Variant Individual with <i>RHDi>*845A/1227A.
Xiu-Hua XIE ; Fan WU ; Qing DENG ; Nai-Bao ZHUANG
Journal of Experimental Hematology 2023;31(4):1150-1154
OBJECTIVE:
To explore the genetic mutation mechanism of a rare Rhesus D variant individual.
METHODS:
Regular serological assay was used for determination of Rh type for the sample. Indirect anti-human globulin test (IAT) was used to confirm the RhD antigen and screen the antibodies. D-screen reagent was used to analyze the RhD epitopes of the sample. <i>RHDi> genotype and <i>RHDi> zygosity testing of the sample were detected by palymerase chain reaction with sequence-specific primers (PCR-SSP). The full length coding region of <i>RHDi> gene was sequenced. <i>RHDi> mRNA was detected using reverse transcription polymerase chain reaction (RT-PCR). The PCR products were cloned and sequenced.
RESULTS:
The RhD blood group of the sample was determined as weak D, and the Rh phenotype was CcDEe. The antibody screening was negative. The sample tested with all monoclonal anti-Ds in D-screen showed the D epitope profiles as partial D types. The analysis of <i>RHDi> gene sequence indicated that the individual with <i>RHDi> c.845G/A and <i>RHDi> c.1227G/A base heterozygosis. Three kinds of alternative splicing isoforms were obtained by TA cloning and sequencing.
CONCLUSION
The object has <i>RHDi> c.845G/A and <i>RHDi> c.1227G/A mutation. This heterozygous mutation is responsible for the low expression of RhD antigen on the red blood cells of the sample.
Alleles
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Blood Group Antigens
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Genotype
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Mutation
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Phenotype
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Polymerase Chain Reaction
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Rh-Hr Blood-Group System/genetics*
;
Humans