1.In-depth identification of para-Bombay blood type in cancer patients using third-generation sequencing technology.
Na WANG ; Xiurong YU ; Yujuan CHEN
Chinese Journal of Cellular and Molecular Immunology 2025;41(2):148-153
Objective To precisely identify the para-Bombay blood types in cancer patients at our hospital, establish a robust system for the identification of challenging blood types in our laboratory, and provide a foundation for precise transfusion practices. Methods We retrospectively analyzed the blood type results of 91 874 cancer patients from January 1, 2019, to December 31, 2023. Conventional serological methods were used to screen for blood types, and suspected para-Bombay blood types were identified. Further analysis was performed using Pacific Biosciences (PacBio) single-molecule real-time sequencing and Sanger sequencing was used to determine the genotypes of the ABO, FUT1, and FUT2 genes. Results Eight cases of para-Bombay blood type were confirmed through serological and molecular biological methods. The FUT1 genotypes identified were: 5 cases of h1h1 (homozygous mutation 551_552delAG) and 3 cases of h1h2 (compound heterozygous mutations of 551_552delAG and 880_882delTT). The FUT2 genotypes identified were: 2 cases of Se357/Se357, 716 and 4 cases of Se357/Se357. Additionally, one sample revealed a novel heterozygous mutation, 818C>T, in exon 7 of the ABO gene, which was confirmed by PacBio sequencing to be located on the O haplotype. Conclusion PacBio sequencing technology demonstrates significant advantages in analyzing the haplotypes of para-Bombay blood type genes. This approach supports the establishment of a robust system for the identification of challenging blood types and provides novel evidence for precise transfusion practices in cancer patients.
Humans
;
Neoplasms/genetics*
;
Fucosyltransferases/genetics*
;
ABO Blood-Group System/genetics*
;
Male
;
High-Throughput Nucleotide Sequencing/methods*
;
Galactoside 2-alpha-L-fucosyltransferase
;
Female
;
Retrospective Studies
;
Genotype
;
Middle Aged
;
Blood Grouping and Crossmatching/methods*
;
Adult
;
Mutation
;
Aged
2.Serological and Molecular Biological Characteristics of cisAB Blood Group and Transfusion Strategies.
Si-Meng WU ; Qiao-Ni YANG ; Wa GAO ; Xiao-Shuai LI ; Qiu-Shi WANG
Journal of Experimental Hematology 2025;33(1):206-210
OBJECTIVE:
To analyze the serological and molecular biological characteristics of 5 patients with cis AB blood group, and to explore the safe transfusion strategy.
METHODS:
Serological identification of the samples' blood group was performed using anti-A, anti-B, anti-D, anti-A1, anti-H typing reagents and ABO reagent erythrocytes. Molecular biological identification of the samples' blood group was performed using PCR-SSP or gene sequencing.
RESULTS:
The serological identification results of blood group in 5 patients all showed inconsistent forward and reverse typing, presenting as A2B3 or A2Bw. ABO gene sequencing of samples 1, 2 and 3 showed 261delG in exon 6 and 467C>T, 803G>C in exon 7. The genotypes of samples 1, 2 and 3 were determined to be cisAB/O . PCR-SSP genotyping was performed on sample 4 and 5,and the results were both cisAB/O .
CONCLUSION
Patients with cisAB alleles have inconsistent serological manifestations, and genetic testing is necessary to ensure the safety and effectiveness of blood transfusion.
Humans
;
ABO Blood-Group System/genetics*
;
Blood Transfusion
;
Blood Grouping and Crossmatching
;
Genotype
;
Blood Group Antigens/genetics*
;
Alleles
;
Male
;
Female
3.A Retrospective Analysis of Irregular Erythrocyte Antibodies in the Blood Transfusion Department of People's Hospital of Xinjiang Uygur Autonomous Region from 2011 to 2022.
Ru-Bin WANG ; Hui-Jun LI ; Fei LI ; Wei CHEN
Journal of Experimental Hematology 2025;33(1):211-216
OBJECTIVE:
The distribution of irregular erythrocyte antibodies in the blood transfusion department of the People's Hospital of Xinjiang Uygur Autonomous Region from 2011 to 2022 and the relationship between irregular erythrocyte antibodies and ethnicity, gender, pregnancy history, blood transfusion history were retrospectively analyzed.
METHODS:
The irregular antibody screening data of patients who were proposed to receive blood transfusions in the clinical blood transfusion safety and blood management software of our hospital from 2011 to 2022 were collected for a retrospective study, and the distribution of irregular erythrocyte antibodies from 2011 to 2022 was analyzed. The relationship between ethnicity, gender, pregnancy history, blood transfusion history and the detection rate of irregular erythrocyte antibodies was further analyzed.
RESULTS:
From 2011 to 2022, the positive detection rate of irregular erythrocyte antibodies in 329 270 samples was 0.77%. Rh blood group (43.72%), Lewis blood group (9.90%) and MNS blood group (6.44%) accounted for the highest proportion of irregular erythrocyte antibody positive samples. In Rh blood group, the proportion of anti-D and anti-E in positive samples was the highest, with 19.09% and 16.06%, respectively. In MNS blood group, the proportion of anti-M in positive samples was the highest (5.46%). In Lewis blood group, the proportion of anti-Lea in positive samples was the highest (8.80%). Compared with other ethnic groups, the detection rates of irregular erythrocyte antibodies were significantly higher in Han, Hui and Uyghur ethnic groups (P < 0.001). Irregular erythrocyte antibody positive samples in Rh blood group system were concentrated in Han and Uygur ethnic groups. Compared to males and patients without a history of blood transfusion and pregnancy, female patients and patients with a history of blood transfusion and pregnancy had significantly higher detection rates of irregulart erythrocyte antibodies (P < 0.01).
CONCLUSION
The results of irregular antibody screening before blood transfusion showed that Rh blood group system antibodies were the main type of irregular antibodies, and the screening of various Rh blood group antigens should be strengthened. And the screening should be focused on female, patients with blood transfusion history and pregnancy history, as well as ethnic minority patients.
Humans
;
Retrospective Studies
;
Female
;
Blood Transfusion
;
China
;
Rh-Hr Blood-Group System/immunology*
;
Male
;
Erythrocytes/immunology*
;
Pregnancy
;
Isoantibodies/blood*
;
Blood Grouping and Crossmatching
;
Antibodies
;
Adult
;
Blood Group Antigens/immunology*
4.Serological and Molecular Biological Detection of RhD Variants.
Dao-Ju REN ; Chun-Yue CHEN ; Xiao-Wei LI ; Jun XIAO ; Xiao-Juan ZHANG ; Cui-Ying LI
Journal of Experimental Hematology 2025;33(2):498-503
OBJECTIVE:
To analyze the RHD genotyping and sequencing results of RhD serology negative samples in the clinic, and to further explore the laboratory methods for RhD detection, in order to provide a basis for clinical precision blood transfusion.
METHODS:
A total of 27 200 whole blood samples were screened for RhD blood group antigen using microcolumn gel card method.Serologic RhD-negative confirmation tests were performed on blood samples that were negative for RhD on initial screening using three different clonal strains of IgG anti-D reagents. The 10 exons of the RHD gene on chromosome 1 were also analyzed by PCR-SSP to determine RHD genotyping.When the PCR-SSP method did not yield definitive results, the RHD gene of the sample was analyzed by the third-generation sequencing.
RESULTS:
The results of the initial screening test by the microcolumn gel card method showed that 136 of the 27 200 samples were RhD-negative, of which 86 underwent RhD-negative confirmation testing and RHD genotyping, 88.37% (76/86 cases) of the RhD-negative confirmation test results were negative for the three anti-D reagents, and the results of RHD genotyping showed that 67.44% (58/86 cases) of the cases had a complete deletion of 10 exons, and the remaining 28 cases were RHD*711delC (1 case), RHD*D-CE(1-9)-D (1 case), RHD*D-CE(2-9-)D (2 cases), RHD*D-CE(3-9)-D (4 cases), RHD*DEL1 (c.1227G >A) mutation (16 cases), RHD*weak partial 15(845G >A) mutation (3 cases), and a mutation of c.165C >T base was found in 1 sample by three-generation sequencing.
CONCLUSION
RHD genotype testing of samples that are serologically negative for RhD antigen shows that some of the samples have RHD gene variants, not all of which are total deletions of RHD, suggesting that there are some limitations of the serologic method for RhD detection. Due to the polymorphism of the RHD gene structure, different RhD variants present different serologic features, which need to be further detected in combination with molecular biology testing, especially for the identification of Asian-type DELs, which is important for clinical precision blood transfusion.
Humans
;
Rh-Hr Blood-Group System/genetics*
;
Genotype
;
Polymerase Chain Reaction
;
Exons
;
Blood Grouping and Crossmatching
5.Analysis of the Influencing Factors of ABO Blood Group Antibody Origin and Titer in Neonates.
Meng-Jiao YANG ; Li ZHANG ; Yu ZHOU ; Chun YANG ; Xiang SHI
Journal of Experimental Hematology 2025;33(2):520-525
OBJECTIVE:
To analyze the origin and influencing factors the titer of ABO blood group antibody in neonates.
METHODS:
A total of 303 newborn blood samples collected in our hospital from August 2023 to March 2024 were selected for the detection of ABO blood group settings and the determination of the total titers of IgG and IgM blood group antibodies in plasma. IgM antibodies were treated with dithithreitol (DTT) to determine the titers of IgG antibodies. The total titer of the blood group antibody was compared with that of the IgG antibody. The clinical data of mothers and newborns were collected, and the correlation between the antibody titer and these clinical data was analyzed.
RESULTS:
Among the 303 newborn specimens, 14 cases (4.62%) were identified to possess blood group antibodies. The influence of the maternal ABO blood group on the generation of high-potency blood group antibodies in newborns was observed to follow the order of O>B>A>AB, with a significant statistical difference ( P < 0.01). Of the 123 (40.59%) newborns born to mothers of type O, 121 (98.37%) had blood group antibody titers > 2. Of the 20 (6.60%) newborns born to mothers of type AB, all 20 (100.00%) had blood group antibody titers < 2. Among 89 (29.37%) mothers of type A and 71 (23.43%) mothers of type B, the titer of 100% newborn blood group antibody was less than 2, when the newborn blood group was incompatible with the mother's blood group; the titer of the newborn blood type antibody was higher or lower, when the newborn blood type was compatible with the mother's blood type. The titer of the newborn blood group antibodies is related to the number of pregnancies of the mothers and has no association with other clinical data (such as the mother's number of obortions), the number of production, fetal gestation age.
CONCLUSION
The majority of ABO blood group antibodies in neonates are IgG antibodies from the mothers, and few are produced by the neonates themselves. In some neonates, IgG anti-A and/or anti-B can agglutinate with anti-stereotyped cells at room temperature. The maternal ABO blood type is the primary factor influencing the titer of the newborn blood type. The number of maternal pregnancies is a factor affecting the high titer ABO blood group antibodies in newborns.
Humans
;
Infant, Newborn
;
ABO Blood-Group System/immunology*
;
Female
;
Immunoglobulin G/blood*
;
Immunoglobulin M/blood*
;
Pregnancy
;
Blood Grouping and Crossmatching
6.Genotyping and Transfusion Strategy for Pregnant Patients with ABO Blood Typing Difficulties.
Chen-Chen FENG ; Qing CHEN ; Xiao WEI ; Li-Li SHI ; Ruo-Yang ZHANG ; Fang ZHAO ; Jian-Yu XIAO
Journal of Experimental Hematology 2025;33(2):538-545
OBJECTIVE:
To identify the blood type of specimens from pregnant patients with difficult-to-type ABO status, and to guide clinical safe blood transfusion.
METHODS:
The specimens from 36 pregnant patients with suspicious ABO blood group were collected. These specimens were submitted by clinical institutions from various regions to our center's genetic testing platform from January 2021 to December 2022. The blood group phenotypes and genotypes of these specimens were identified by serological method and genetic sequencing.
RESULTS:
A total of 20 ABO subtypes were detected in the 36 samples, including 10 cases of BA/O, 3 cases of cisAB/O, 2 cases of A/Bw, 1 case of A2/B, 1 case of Aw/B, 1 case of BA/B, 1 case of BA/A, and 1 case of Bw/O. Additionally, 4 cases were identified as para-Bombay blood type, and no specific variations associated with abnormal phenotypes were found in the remaining 12 cases.
CONCLUSION
ABO subtypes interfere with ABO blood group identification in pregnant patients, and pregnancy status also affects blood group phenotype. Accurate determination of blood group genotype by genetic sequencing technology can guide clinical blood transfusion for pregnant patients, and ensure maternal and infant safety.
Humans
;
Female
;
Pregnancy
;
ABO Blood-Group System/genetics*
;
Blood Grouping and Crossmatching
;
Blood Transfusion
;
Genotype
;
Phenotype
7.Analysis of Positive Results of Anti-M Unexpected Antibody in Pediatric Inpatients in Central China.
Dong-Dong TIAN ; Ding ZHAO ; Wei LI ; Yong-Jun WANG ; Hong-Bing HU ; Yuan-Qing YANG ; Zheng-Feng LI
Journal of Experimental Hematology 2025;33(4):1155-1160
OBJECTIVE:
To analyze the positive rate and distribution of anti-M unexpected antibody in pediatric inpatients aged 0 to 14 years in central China.
METHODS:
A total of 30 049 pediatric inpatients admitted to the Second Xiangya Hospital of Central South University, Wuhan Children's Hospital and Children's Hospital Affiliated of Zhengzhou University from May 2020 to August 2022 were enrolled in this study, and relevant clinical data were collected. Blood samples from the patients were tested for blood typing and screened for unexpected antibodies. For samples that screened positive for unexpected antibodies, identification was conducted using the identification panel to determine the specificity of the antibodies. The distribution and differences of anti-M antibodies in pediatric patients of different sexes, ages, blood groups, disease types, with or without a history of blood transfusion, and across different regions were analyzed.
RESULTS:
Among 30 049 inpatients, the positive rate of unexpected antibodies was 0.91% (273/30 049), of which the positive rate of anti-M antibodies was 0.44% (131/30 049). The positive rate of anti-M antibodies in the neonates aged 0 to < 1 month was 0.10% (5/4 881), and all of them were IgG antibodies from their mothers; The positive rate of anti-M antibodies for the group aged from 1 month to < 1 year old was 0.23% (7/3 108), with no anti-M antibodies detected in patients aged 1-6 months; The positive rates of anti-M antibodies in the 1-4 years old group, 5-9 years old group, and 10-14 years old group were 0.87% (88/10 064), 0.38% (27/7 190), and 0.08% (4/4 806), respectively. The positive rate of anti-M antibodies in the 1-4 years old group was significantly higher than that of the other groups ( P <0.001), and there were also statistical differences in the positive rate between the 5-9 years old group and the 0-< 1 month and 10-14 years old groups ( P <0.001). The prevalences of anti-M antibodies in ABO blood group A, B, O and AB were 0.32% (30/9 482), 0.70% (58/8 293), 0.32% (31/9 595) and 0.45% (12/2 679), respectively. The prevalence of anti-M antibodies in patients with blood group B was significantly higher than that in patients with blood groups A and O ( P <0.05). The prevalences of anti-M antibodies in Hunan, Hubei and Henan was 0.18%, 0.32% and 0.71%, respectively. The prevalence of anti-M antibodies in Henan was significantly higher than that in Hunan and Hubei ( P <0.05), and the distribution showed obvious regional differences between the north and the south. There were no significant differences in the positive rate of anti-M antibodies between the children with different sexes, disease types, and with or without a history of blood transfusion (P >0.05).
CONCLUSION
This study reveals the distribution pattern of anti-M antibodies in pediatric inpatients aged 0-14 years in central China, which has reference value for the research on unexpected red blood cell antibodies in Chinese children.
Humans
;
Child
;
China
;
Infant
;
Child, Preschool
;
Adolescent
;
Female
;
Male
;
Inpatients
;
Infant, Newborn
;
Blood Grouping and Crossmatching
;
Antibodies/blood*
8.Detection and Transfusion Strategy of Mimicking Antibodies.
Hui ZHANG ; Jie-Wei ZHENG ; Sha JIN ; Wei SHEN ; Shan-Shan LI ; Xiao-Wen CHENG ; Dong XIANG
Journal of Experimental Hematology 2025;33(4):1168-1172
OBJECTIVE:
To explore serological detection and blood transfusion strategies of mimicking antibodies, so as to provide appropriate transfusion strategies.
METHODS:
Detailed serological tests, including ABO blood group, Rh typing, antibody specificity, etc,were performed on two patients with autoimmune hemolytic anemia(AIHA). Meanwhile, the references about blood transfusion from mimicking antibody patients published from 1977 to 2024 in China and abroad were retrospectively summarized and analyzed.
RESULTS:
The patient 1 blood type was AB,CCDee and the antibody is mimicking anti-e, transfusion the e-negative red blood cells (RBCs) was effective. After two transfusions of e-RBCs, hemoglobin levels significantly increased from 48 g/L to 91 g/L, with complete resolution of hemolytic symptoms. The patient 2 blood type was O,CcDee, and the antibody was mimicking anti-c, the patient was diagnosed with AIHA and treated with hormone. No blood products were transfused during hospitalization, and his hemolysis was relieved.
CONCLUSION
Strictly grasping the indication of blood transfusion, blood transfusion should not be performed in the unnecessary conditions, and the corresponding antigen-negative RBC should be screened for transfusion in the necessay conditions.
Humans
;
Blood Transfusion
;
Anemia, Hemolytic, Autoimmune/therapy*
;
ABO Blood-Group System
;
Retrospective Studies
;
Antibodies
;
Male
;
Blood Grouping and Crossmatching
9.Serological and Molecular Biological Analysis of a B(A) Subtype Family and Strategies for Safe Blood Transfusion.
Ni-Na WANG ; Hong-Hong ZHANG ; Fu-Ting SUN ; Jun SU
Journal of Experimental Hematology 2025;33(5):1412-1417
OBJECTIVE:
Serological and molecular biological analysis of a B(A) subtype family was carried out to explore the underlying mechanism of B(A) subtype and clinical safe blood transfusion strategies.
METHODS:
The ABO blood type of the proband and her four family members were identified by serological methods, and serological experiments such as anti-H, anti-A1 and absorption-elution tests was added. In addition, the exons 6 and 7 of the ABO gene were sequenced by PCR-SSP (polymerase chain reaction - sequence specific primer).
RESULTS:
The serological results showed that the agglutination intensity of the proband, her mother and her maternal grandmother was imbalanced during forward typing, showing weak A and strong B antigens, and there were strong H antigens and their intensity were higher than that of normal B type. The results of reverse typing indicated the presence of weak anti-A1 antibodies, and human anti-A was positive in the absorption-elution test. Genetic sequencing revealed a characteristic mutation of c.700 C>G in all three individuals. The sequencing results showed that the proband was B(A)02/B01, her mother was B(A)02/O02, and her maternal grandmother was B(A)02/O01 . According to the compatibility principle, 1.5 units of type O washed red blood cells were transfused intraoperatively, resulting in no adverse reactions.
CONCLUSION
The c.700 C > G mutation on exon 7 is the molecular basis for the formation of B(A)02, and pedigree analysis shows that the B(A)02 allele was inherited from the proband's maternal grandmother to the proband's mother and then to the proband, showing a stable cis-inheritance pattern rather than a spontaneous mutation. For patients with B(A)02 subtype, type O washed red blood cells and type AB plasma can be transfused according to the principle of compatibility.
Humans
;
ABO Blood-Group System/genetics*
;
Female
;
Blood Transfusion
;
Blood Grouping and Crossmatching
;
Pedigree
;
Male
;
Mutation
;
Adult
;
Exons
10.Establishment and Preliminary Application of qPCR-Based Genotyping Method for Diego, MNS and Kell Blood Groups of Red Blood Cells.
Bing ZHANG ; Gang XU ; Wen-Jian HU ; Xiao-Zhen HONG ; Xian-Guo XU
Journal of Experimental Hematology 2025;33(5):1429-1434
OBJECTIVE:
To establish a genotyping method for Diego, MNS and Kell blood groups based on quantitative real-time PCR (qPCR) technology, and preliminarily apply it to the screening of rare blood groups in blood donors.
METHODS:
Blood group gene standards containing heterozygous and homozygous alleles were prepared by blood group serological and PCR-SBT methods. Specific amplification primers and hybridization probes were designed, and explore to establish the qPCR method for detecting Diego, MNS, and Kell blood group genotypes. Then the established qPCR method was used to identify blood group genotypes of 186 blood donor samples.
RESULTS:
A method based on qPCR technology was established to identify Dia/Dib, S/s and K/k blood group antigens. The genotyping results of the gene standard samples were consistent with the serological testing results and genotypes detected by PCR-SBT. qPCR testing of 186 samples identified 11 cases of DI*A/B heterozygosity and 19 cases of GYPB*S/s heterozygosity, and the rest were DI*B/B, GYPB*s/s, KEL*02/02 homozygosity. No rare blood group genotypes of DI*A/A, GYPB*S/S, KEL*01.01/01.01 were found.
CONCLUSION
The established qPCR method is suitable for genotyping on Diego, MNS and Kell blood group, and it can be used for batch screening of blood donors and the establishment of rare blood group bank.
Humans
;
Genotype
;
Genotyping Techniques/methods*
;
Real-Time Polymerase Chain Reaction/methods*
;
Blood Group Antigens/genetics*
;
Kell Blood-Group System/genetics*
;
Blood Donors
;
Blood Grouping and Crossmatching/methods*
;
Erythrocytes
;
MNSs Blood-Group System/genetics*

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