Genotyping of the rare Para-Bombay blood group in southern Thailand

ABSTRACT Introduction: The para-Bombay phenotype, or H-deficient secretor, results from different mutations of the FUT1, with or without the FUT2 mutation. Consequently, there is an absent or weak expression of the H antigen on red blood cells (RBCs). Routine ABO blood grouping for two siblings with blood group O showed discrepant results with their parental blood group AB. Fragments encompassing the entire coding region of the FUT1 and FUT2 genes were investigated. Methods: Blood and saliva specimens were collected to verify the correct ABO grouping by cell grouping, serum grouping and the hemagglutination inhibition (HI) test, respectively. The FUT1 and FUT2 genomes were identified using the whole-exome sequencing (WES) in two children's DNA blood specimens and may have caused, or been relative to, their blood group. Genetic variations of the FUT1 and FUT2 genes have been investigated in the other family members using the Sanger sequencing. Results: The serologic reaction results of the proband revealed that A, B and H antigens were absent on RBCs, and that the serum contained anti-H. However, ABH and AH antigens were present in the saliva PB1 and PB2, respectively. The probands PB1 and PB2 were assigned as AB and A blood groups, respectively. Blood genotyping confirmed that heterozygous mutations of the FUT1 gene, c.551_552delAG, were identified. Three family members, PB3, PB, and PB8, also showed normal ABO blood groups, but their genotypes were also the FUT1 mutation c.551_552delAG. Conclusions: The FUT1 mutation c.551_552delAG may result in the reduced or absent H antigen production on RBCs, which characterizes the para-Bombay phenotypes. Blood genotyping is essential if these individuals need a blood transfusion or are planning to donate blood.

Comparison of ABO Blood Group Typing between Automated Blood Bank Analyzer IH-500 and Manual Method / 대한수혈학회지

BACKGROUND: According to increased availability and awareness of automated blood bank analyzer with its speed and efficiency, use of automated analyzer in hospital blood bank has been increasing rapidly. We compared the ABO blood group typing results between automated analyzer IH-500 and manual method in healthy adults and patients with ABO discrepancies to provide useful information on interpretation of blood grouping results by automated analyzer. METHODS: Among healthy adults who underwent medical examinations, 400 samples (each 100 samples of A, B, O and AB type) were selected and evaluated the results and grades of blood grouping by automated and manual methods. Also, 50 samples showing ABO discrepancies among patients requested for pretransfusion test were selected and compared between two methods. As for samples with ABO discrepancies, further tests such as microscopic examination, reactivity with anti-A1 or ABO genotyping along with medical record review were performed. RESULTS: Agglutination results and grades in healthy adults were consistent between two methods. Meanwhile, 30 (60%) of ABO discrepant samples were related to rouleaux formation and their frequencies and agglutination grades were higher in automated method (Wilcoxon signed rank test, P=0.001). Results of discrepant samples caused by unexpected antibody or ABO subgroup showed no differences between two methods. CONCLUSION: IH-500 automated analyzer was considered useful for mass examination of healthy individuals. Meanwhile, considering the fact that ABO discrepancies by rouleaux formation were more frequent and stronger in automated method, it is recommended to retest their results by manual methods along with medical record review.

Analysis of ABO Blood Discrepancies and Transfusion Experiences in Chonnam National University Hospital / 대한수혈학회지

BACKGROUND: ABO blood group discrepancy occurs when the results of red cell tests do not agree with those of the serum test. In order to select the proper blood units for transfusion, clarification of the cause of ABO discrepancies is essential. We analyzed the cases and recent actual transfusion experiences at Chonnam National University Hospital (CNUH). METHODS: In total, among pre-transfusion blood samples at CNUH between January 2012 and July 2013, 55 cases of ABO discrepancies were analyzed retrospectively. RESULTS: The discrepancy incidence was 0.14%. Problems with serum were the most common cause of ABO discrepancies, with 31 cases (56.4%), and extra serum reactivity due to cold allo-antibodies accounted for the highest frequency (n=7). There were three cases of non-specific aggregations caused by commercial RBC constituents and aggregation was not observed when a re-test was performed with other commercial RBCs or self-prepared human RBCs. Two of three cases with mix-field aggregations involved a pair of twins after in vitro fertilization - embryo transfer (IVF-ET). Among 55 patients, 20 were actually transfused, and all but four cases had weaker or identical RBC units and stronger or identical plasma units. CONCLUSION: There were newly revealed ABO discrepancies caused by non-specific aggregations of commercial RBCs and in twins after IVF-ET. In addition, investigation of actual transfusion experiences in patients with ABO blood group discrepancies would be helpful.

A Case of IgM Deficiency with B Cell Deficiency Detected by ABO Discrepancy in a Patient with Acute Osteomyelitis

ABO discrepancy refers to an inconsistency between red cell and serum typings and has various causes, including hypogammaglobulinemia. IgM deficiency is a rare disorder that may accompany several conditions such as infection and autoimmune disorders. Here, we describe a case of IgM deficiency discovered during the evaluation of an ABO discrepancy in a 16-yr-old Korean boy. ABO blood grouping showed that while his cell type was O+, serum typing detected only anti-A (3+). Anti-B was not detectable at room temperature but was graded at 1+ at 4degrees C. ABO genotyping revealed an O/O genotype. His serum IgG, IgA, and IgM concentrations were 770 mg/dL (reference range: 800-1,700 mg/dL), 244 mg/dL (reference range: 100-490 mg/dL), and 13.5 mg/dL (reference range: 50-320 mg/dL), respectively. He was diagnosed with acute osteomyelitis on the basis of clinical presentation and imaging studies. The symptoms gradually improved within 3 weeks of treatment. However, the ABO discrepancy and IgM deficiency persisted even 6 months after recovery and lymphocyte subset analysis revealed CD19+ B cell deficiency. To the best of our knowledge, IgM deficiency detected by ABO discrepancy in a patient with acute osteomyelitis has not been reported before.

A Case of IgM Deficiency with B Cell Deficiency Detected by ABO Discrepancy in a Patient with Acute Osteomyelitis

ABO discrepancy refers to an inconsistency between red cell and serum typings and has various causes, including hypogammaglobulinemia. IgM deficiency is a rare disorder that may accompany several conditions such as infection and autoimmune disorders. Here, we describe a case of IgM deficiency discovered during the evaluation of an ABO discrepancy in a 16-yr-old Korean boy. ABO blood grouping showed that while his cell type was O+, serum typing detected only anti-A (3+). Anti-B was not detectable at room temperature but was graded at 1+ at 4degrees C. ABO genotyping revealed an O/O genotype. His serum IgG, IgA, and IgM concentrations were 770 mg/dL (reference range: 800-1,700 mg/dL), 244 mg/dL (reference range: 100-490 mg/dL), and 13.5 mg/dL (reference range: 50-320 mg/dL), respectively. He was diagnosed with acute osteomyelitis on the basis of clinical presentation and imaging studies. The symptoms gradually improved within 3 weeks of treatment. However, the ABO discrepancy and IgM deficiency persisted even 6 months after recovery and lymphocyte subset analysis revealed CD19+ B cell deficiency. To the best of our knowledge, IgM deficiency detected by ABO discrepancy in a patient with acute osteomyelitis has not been reported before.

Naturally occurring anti M complicating ABO grouping.

Anti M is considered a naturally occurring antibody that is usually active at temperatures below 37˚C and is thus of on clinical significance. This antibody, if present in an individual, can lead to a discrepancy between forward and reverse ABO grouping and thus creates diagnostic difficulties for blood bank staff. We report a case of a 58-year-old lady who had an unexpected reaction in reverse grouping due to anti M that posed a problem for us in the significance of such discrepancy in blood grouping.

New Variant of O02 Allele Found in the Family of Cis-AB: Report of Three Cases in a Family / 대한수혈학회지

In preoperative ABO typing, we observed an ABO discrepancy in a 61 year-old patient with A2B3 phenotype. Standard serologic tests for the ABO blood group phenotypes and ABO gene direct sequencing for the exons 6 and 7 were done as part of the patient's family study. We found that the patient's genotype was identical to cis-AB01/O02, except for a 703 G>A polymorphism at exon 7. An allele-separation by cloning and subsequent sequencing of the heterozygote was carried out. We identified a novel O02 variant allele characterized by a 703 G>A polymorphism in the patient and in her 2 daughters.

Chimerism Proven by Flow Cytomety in the Setting of an ABO Discrepancy: A Case Report / 대한수혈학회지

Chimerism is an important, yet uncommon cause of ABO phenotype/genotype discrepancies. The propositus was a 28 year-old pregnant women who had an ABO discrepancy, expressing an A(weak)B RBC phenotype and AB reverse type. Sequencing of ABO exons 6 and 7 revealed a B101/O01 genotype. Neither analysis of 9 short tandem repeats (STR) loci nor HLA typing on DNA extracted from white blood cells demonstrated evidence of chimerism. However, flow cytometric analysis using a PE-conjugated anti-A mouse monoclonal antibody detected a small population (3.8%) of red cells expressing normal A antigen. Based on this, we suggest that flow cytometric analysis is an effective method for the identification of small chimeric populations of RBCs in the immunohematology laboratory.

A Case of Group O Losing Anti-B Selectively / 대한수혈학회지

We report a case of group O losing anti-B selectively. A 25-year-old male donated blood; on the donor test an ABO discrepancy was noted, and a further evaluation study was performed. ABO genotyping with an allele specific polymerase chain reaction assay revealed O/O and DNA sequencing of exons 6 and 7 of the ABO gene showed O01/O02. The serum gammaglobulin level was decreased and only 0.2% CD19 pan-B positive lymphocytes were present in a subset of lymphocytes. In a previous donor study, anti-B of the patient was lost from a third donor study and was still not detected.

A Case of ABO Discrepancy due to Anti-IH / 대한수혈학회지

The patient was a 70-year-old woman with hypertension and end stage renal disease, and she presented with left wrist pain due to falling a day before admission. On admission, laboratory testing revealed a hemoglobin level of 6.7 g/dL, and a physician ordered 2 units of packed RBCs. She had never received a RBC transfusion in the past. The ABO grouping showed a discrepancy between the cell type AB and serum type O, and the irregular antibody screening was negative. Crossmatchings with group AB and group O RBCs were incompatible. Anti-I, which is a cold antibody, was inferred because the degree of agglutination was decreased after warming. However, crossmatching with group O RBCs, which are the universal donor blood, was positive and the anti-IH was considered to be the specificity of the irregular antibody. The patient's serum did not react with group O cord (i) blood cells and anti-I was then considered. The genotype of this patient was AB, and it was inferred that the ABO discrepancy was due to anti-IH.

ABO Gene Analysis of Discrepant ABO Blood Group in Blood Donors / 대한수혈학회지

BACKGROUND: An exact ABO blood group is essential for prevention of transfusion accident and safe transfusion therapy. It is known that one of causes of ABO discrepancies is ABO subgroup caused by genetic polymorphism. Therefore, we analyzed ABO genotype of ABO discrepancies in blood donors and studied the distribution and cause of ABO discrepancies. METHODS: This study examined 118 samples showing ABO discrepancies of ABO blood typing between May 2003 and Dec 2003. ABO genotyping using the polymerase chain reaction-restriction fragment length polymorphism(PCR-RFLP) method was performed on 118 samples. Restriction enzymes including BssH II, Kpn I and Alu I were used for PCR-RFLP. RESULTS: The genotypes of 118 cases were composed of 43 cases of A/B, 12 cases of A/O, 10 cases of B/O, 1 case of B/B, 37 cases of cis-AB/O, 4 cases of cis-AB/A, 11 cases of cis-AB/B. The genotype of cis-AB/O showed 32 cases with phenotype A2 B3 , 2 cases with phenotype A2 B, 2 cases with phenotype A1 B3 , 1 case with phenotype Ael B. The genotype of cis-AB/B showed 11 cases with phenotype A2 B, and cis-AB/A showed 2 cases with phenotype A2 B3 , 1 case with phenotype A1 Bx and 1 case with phenotype A1 Bel. CONCLUSION: These data demonstrated that the most frequent genotype of ABO discrepancies in our study is cis-AB. The most predominent phenotype of cis-AB/O is A2 B3 . ABO genotyping is useful in resolving ABO discrepancies, and determination of ABO subgroups.

A Case of Mixed-type Autoimmune Hemolytic Anemia / 대한혈액학회지

We report a case of mixed-type autoimmune hemolytic anemia in a 73-year-old female. At first, she visited at Pusan National University Hospital, complaining of dizziness and dyspnea for 3 months. Her hemoglobin level was 5.5g/dL, so red blood transfusion was requested by her physician. In ABO blood grouping, a discrepancy between cell and serum typing was observed. Also all of donor cells for crossmatching were reactive with patient's serum. High mean corpuscular volume (MCV) value was not normalized after warming. The cold agglutinin titer was 1 : 64 at 4degrees C, and was increased up to 1 : 1,024 at room temperature and 37degrees C, suggesting mixed-type autoimmune hemolytic anemia. The patient's symptoms and signs were improved progressively with corticosteroid therapy, and also ABO discrepancy was disappeared.

Two Cases of ABO Discrepancy due to Hypogammaglobulinemia / 대한수혈학회지

Two cases of ABO discrepancy were observed in thirty-year old woman with gall bladder abscess and fifty-five-year old woman with hepatocellular carcinoma. Their red cells were typed as group O and their serum had only anti-A antibody. Absence of A and B antigens on their RBCs were confirmed by adsorption elution test and saliva test. The B transferase activities were not demonstrated in their serum. Their ABO genotypes were O/O by sequence specific polymerase chain reaction. Their serum protein electrophoresis showed hypogammaglobulinemia pattern, and immunoglobulin levels (IgG, IgA, IgM) were decreased (39 mg/dL, 46 mg/dL, <5 mg/dL and 63 mg/dL, 65 mg/dL, 12 mg/dL, respectively).

A Case of Agammaglobulinemia Detected by ABO Discrepancy in a 13-year-old Girl / 대한진단검사의학회지

We report a case of agammaglobulinemia detected by ABO discrepancy in a 13-year-old girl. At first, she visited Pusan National University Hospital complaining of joint swelling and pain. For trans-fusion, the ABO blood grouping was done. The cell typing was B, but anti-A was not found in the serum with repeated testing. Then, serum protein electrophoresis, immunoelectrophoresis and immunoglob-ulin quantitation showed a markedly decreased level in gamma-globulin. B lymphocyte was less than 1% of peripheral lymphocytes. There were no remarkable abnormal findings in the clinical and laboratory results of her parents. The patient represented an apparent agammaglobulinemia without clinically significant infection for three months. Further follow-up is needed to evaluate the patient.

A Case of BX / 대한수혈학회지

We report a case of BX in a 27-year-old woman admitted for Cessarian section after diagnosis of placenta previa. An ABO discrepancy was observed, such as, group O in cell typing but group B in serum typing. The patient's red cells were agglutinated by anti-H and show the mixed field agglutination by anti-A,B. The results of adsorption and elution test, saliva test and B transferase test show the presence of B antigen, absence of B substance in saliva and no activity of B-transferase. All of these findings were compatible with BX.

Application of ABO genotyping in determination of ABO subgroups / 대한수혈학회지

BACKGROUND: The knowledge about the nucleotides sequence of 9th chromosome that regulates the phenotype of ABO blood group has made the ABO genotyping possible. Since the genotyping can be done with only a small amount of DNA sample, it was primarily applied to the field of forensic medicine. When applied to the blood bank, it is useful in the resolution for ABO discrepancies between the cell and serum typing and determination of A and B subgroups. Rapid ABO genotyping using the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method and its value in determination of ABO subgroups is presented. METHODS: ABO genotyping was performed in seven patients and three families, seven were the cases of ABO discrepancies in routine ABO grouping and three families were for the confirmation of the ABO group. To identify the 261th nucleotide, a 252 bp PCR amplifed fragment was amplified by PCR and digested with Kpn I. For 703th nucleotide, a 128 bp PCR amplified fragment was designed and digested with Alu I. To determine the ABO genotype, the patterns of digestion in DNA fragment were examined. RESULTS: Among the seven cases of ABO discrepancies, B3 and Ael were two cases each. Weakened B due to leukemia was the one, and the other two cases were cis-AB and Am. The three families for confirmation of the ABO group were acquired B due to infection one family, cis-AB two families. CONCLUSIONS: ABO genotyping is a rapid and reliable method that can be used in the case of ABO discrepancies and determination of ABO subgroups.

Three cases of multiple myeloma showing ABO discrepancy / 대한수혈학회지

We report 3 cases of multiple myeloma showing ABO discrepancy with missed reaction in serum typing. They showed markedly decreased immunogolobulin level except for monoclonally increased abnormal immunoglobulin. Their blood group was confirmed by saliva test and addition of anti-globulin reagent. As serum immunoglobulin level is raised, the reactivity in serum typing showed improving tendency and ABO discrepancy appeared when immunoglobulin was markedly decreased.