Molecular genetic analysis and predigree of a new FUT1 allele of para-Bombay blood group: a case report / 中国输血杂志

【Objective】 To study the serological and molecular mechanism of a case of para-Bombay blood group caused by 236delG mutation of FUT1 gene and investigate the pedigree. 【Methods】 The ABO, H and Lewis antigens of the proband and her family members were detected serologically, and the ABO blood group was confirmed by gene testing. The FUT1 gene was amplified by PCR and then sequenced. The structure of FUT1 236delG enzyme of the proband was simulated in 3D by SwissModel online server. 【Results】 Serological results showed that the proband was rare para-Bombay ABhm, Le(a-b-). Her father and mother was type A and type B, respectively. The gene results showed that the proband was type AB, while her father and mother was type A and type B, respectively. The sequencing results showed that the proband had 236delG/551_552delAG gene mutation, while her mother had 236delG FUT1 gene mutation, and her father had 551_552delAG FUT1 gene mutation. The 3D simulation of the enzyme structure of the proband FUT1 236delG showed that the translated product was an alpha helix structure with no actual function. 【Conclusion】 The 236delG mutation is a new discovered mutation in FUT1 genotype, with 551_ 552delAG mutation(FUT1* 01N.06 genotype), which can result in the generation of para-Bombay blood group.

Para-Bombay phenotype identification and transfusion strategy / 中国输血杂志

【Objective】 To investigate the serological and molecular biological characteristics of para-Bombay phenotype. 【Methods】 ABO blood type, H antigen and Lewis blood type in one blood sample with discrepancy between forward and reverse blood typing were detected by serological method. Antibody screening and identification and cross-match test were also performed by serological method. ABO blood group genes were detected by PCR-SSP, and FUT1 and FUT2 genes were sequenced. 【Results】 The serological test showed that the Para-Bombay phenotype was Ah secretion. The ABO blood group gene was AO2. FUT1 sequencing revealed two mutations: 235G>C and 881_882delTT. While FUT2 sequencing showed only one mutation 357C>T. 【Conclusion】 The discovery and accurate identification of blood groups is necessary to ensure the safety of blood transfusion. Blood donors of rare blood groups should be informed and recruited to the team of rare blood donors.

Study on molecular mechanism and pedigree investigationof para-bombay blood group caused by rare mutations at position h235 and position h649 of FUT1 gene / 中国输血杂志

【Objective】 To study the molecular mechanism of para-bombay blood group caused by two rare mutation combinations and investigate the pedigree. 【Methods】 ABO and H antigens were detected by serological test, and the ABO blood group was confirmed by gene detection; the FUT1 gene was amplified by PCR and sequenced. 【Results】 Serological results showed that the proband was a rare para-bombay blood Bhm, Le(a-b+ ), and the proband′s parents were both B Le (a-b + ). The sequencing results showed that the proband and his mother had mutation at h235 G→C of FUT1 gene. Proband and his father had mutation at h649 G→T of FUT1 gene. 【Conclusion】 The combination of two rare mutations resulted in the formation of a para-bombay phenotype, which is helpful to clarify the gene polymorphism and genetic heterogeneity of para-bombay phenotype, providing data support and theoretical basis for its blood group identification and safe and reasonable blood use of para-bombay phenotype.

Bombay Blood Group– Detection And Study Of Prevalence Of This Rare Phenotype Along With The Transfusion Recommendations

Four main blood types routinely identified are A,B,AB and O. Bombay phenotype individuals are typed as group O on forward ABO typing. Their red cells lack A,B,H antigens and their sera contain anti-A, anti-B and anti-H. It is important to correctly type individuals who are Bombay phenotypes because these individuals require autologous blood donation or blood from another Bombay individual. The present prospective study was conducted over two years to study the prevalence of Bombay phenotype with transfusion recommendations to the blood recipients. All the donor and patient’s blood group were confirmed by tube method. All blood samples showing O blood group on froward grouping and agglutination with O cell in reverse grouping, were tested for Bombay blood group using anti-H.Out of 76,204 cases constituting 49,604 donors and 26,600 patients, Bombay phenotype was detected in 12 cases (0.015%) constituting 4 number of donors and 8 number of patients. All cases were further ruled out to be para Bombay phenotypes and were found to be non secretor by agglutination inhibition test. Four cases out of 12 patients requiring blood transfusion 3 could be issued Bombay blood group and but death occurred in one case due to delay in the surgery for the unavailability of this rare phenotype. Thus, it is recommended that all blood group donors and patients should be routinely screened by both forward and reverse grouping for screening of Bombay phenotype to reduce the risk of hemolytic transfusion reaction resulting from issue of O blood group to Bombay blood group recipients

Identification of Salmonella H antigens by xTAG technology / 中华微生物学和免疫学杂志

Objective To develop a microsphere-based suspension array for simultaneous detec-tion and identification of Salmonella H antigens by using Luminex xTAG technology and to evaluate its capa-bility in serotyping Salmonella strains. Methods The fliC and fljB genes, encoding the H antigen of Salmo-nella, were selected as the target genes. Universal upstream primers were designed based on the highly con-served regions of fliC and fljB genes, and the corresponding specific reverse primers were designed based on the variable regions. While synthesizing, the 5′end of each upstream primer was labeled with biotin and the 5′end of each specific reverse primer was modified with its certain TAG sequence. After amplified and la-beled with biotin and TAG sequence, the PCR products of specimens were hybridized with the mixture of va-rious MagPlex-xTAGTM microspheres. Each set of microspheres contained its unique anti-TAG sequences. The results of hybridization were analyzed by using Luminex MagPix reader system and the median fluores-cence intensity ( MFI) was reported. The H antigens of 145 Salmonella strains were identified with this de-veloped xTAG suspension array, and the results were compared with those obtained by using traditional ser-um agglutination test. Results The PCR products of different H antigens ranged from 94 bp to 245 bp and could be identified by hybridizing with MagPlex-xTAGTM microspheres. There was no cross-reaction between different H antigens or with DNAs derived from Escherichia coli, Vibrio cholerae, Vibrio parahaemolyticus and Shigella flexneri. Compared with the traditional serum agglutination test, the sensitivity and specificity of the xTAG suspension array in the identification of H antigens of 145 Salmonella strains were 95. 1% and 100%, respectively. Conclusion The developed xTAG suspension array was a specific, accurate and effective method for simultaneous detection and identification of 31 H antigens of common Salmonella serovars strains. It could be used for determining the H antigens of more than 90 Salmonella strains within 5 hours.

Identification of new flagellin-encoding fliC genes in Escherichia coli isolated from domestic animals using RFLP-PCR and sequencing methods / Identificação de novas flagelinas codificadas por fliC em Escherichia coli isoladas de animais domésticos utilizando RFLP-PCR e sequenciamento

Identification of Escherichia coli requires knowledge regarding the prevalent serotypes and virulence factors profiles allows the classification in pathogenic/non-pathogenic. However, some of these bacteria do not express flagellar antigen invitro. In this case the PCR-restriction fragment length polymorphism (RFLP-PCR) and sequencing of the fliC may be suitable for the identification of antigens by replacing the traditional serology. We studied 17 samples of E. coli isolated from animals and presenting antigen H nontypeable (HNT). The H antigens were characterized by PCR-RFLP and sequencing of fliC gene. Three new flagellin genes were identified, for which specific antisera were obtained. The PCR-RFLP was shown to be faster than the serotyping H antigen in E. coli, provided information on some characteristics of these antigens and indicated the presence of new genes fliC.

A identificação da Escherichia coli requer conhecimento sobre os sorotipos e fatores de virulência prevalentes permitindo a classificação em patogênico/não patogênico. No entanto, algumas destas bactérias não expressam o antígeno flagelar in vitro. Neste caso, o PCR-restriction fragment length polymorphism (RFLP-PCR) e o sequenciamento do gene fliC podem ser adequados para a identificação desses antígenos, substituindo a sorologia tradicional. Nesta pesquisa foram estudadas 17 amostras de E. coli isoladas de animais e que apresentavam antígeno H não tipável (HNT). Os antígenos H foram caracterizados por PCR-RFLP e sequenciamento do gene fliC. Três novos genes da flagelina foram identificados, para os quais anti-soros específicos foram obtidos. A técnica PCR-RFLP mostrou-se mais rápida que a sorotipagem do antígeno H em E. coli, fornecendo informações sobre algumas características desses antígenos e indicou a presença de novos genes fliC.

Investigation of H antigen on AB subgroup RBCs / 中国输血杂志

Objective To find the rule of the distribution of H antigens on AB subgrouperythrocytes.Methods ABO subgroups were confirmed by using serological and molecular biology (PCR-RFLP) methods. AB subgroup with strong H was defined as red cell agglutination by anti-H of 2 scores or more higher than that of B cells.Results Strong H was only found in certain AB subgroups ,CisAB(100%),B(A)(100%), AxB(46.2%) and A2B(43.6%), but seldom in others among Chinese population.Conclusion The fact that H type-3, which comes from A type-2, can hardly be transferred by B and weak A glycosyltransferase can help to explain why some ‘strong’ H combines with ‘weak’ A in AB erythrocytes. Why only little H can be found in 53.8% AxB, 56.4% A2B and all A3B, AmB subgroup samples still cannot been explained.