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【Objective】 To determine molecular basis of a rare HLA-A typing results carrying triple A alleles in potential allo-HSCT donor and her family. 【Methods】 HLA-A, -B, -C, -DRB1, -DQB1, -E, -F, -G of 5 members in the family were genotyped at a high-resolution level using next-generation sequencing (NGS). HLA-A of probosita was re-checked using polymerase chain reaction-sequence-based typing (PCR-SBT), and SNP oligonucleotide probes (SNP-array)were scanned with genomic DNA of probosita. 【Results】 There was 162.9Kb duplication in 6p22.1(29, 803, 377-29, 966, 301)of probosita who carried triple A alleles A*02∶01∶01, A*11∶01∶01, A*24∶02∶01. Other two family members were found to carry this haplotype: A*02∶01∶01, A*24∶02∶01, B*54∶01∶01, C*01∶02∶01, DRB1*04∶05∶01, DQB1*04∶01∶01, E*01∶01∶01∶03, F*01∶01∶01, G*01∶01∶01∶01, which as a Mendelian gene was segregated and stably transmitted through two generations. 【Conclusion】 Tiny gene duplication induces one haplotype carries two HLA-A alleles in a potential healthy donor for allo-transplantaion and stably transmits through two generations.Routine HLA typing laboratories should pay more attention to this situation and accurately report.
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【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.
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【Objective】 To investigate the serological and molecular genetic characteristics of B(A) and cisAB blood groups discovered in our laboratory. 【Methods】 ABO blood group serology and genetic tests were used to identify blood groups of 6 blood samples, submitted by blood center and hospitals in Shandong, and pedigree investigation was carried on 2 of them. 【Results】 Among the 6 samples, serological results were B(A) in 5 cases and cisAB in 1 case. The results of genetic tests were consistent with the serological results, as the alleles included B(A)04, B(A)02 and cisAB01, and all genotypes were heterozygous with O. Serological pedigree study was conducted on 2 samples: One cisAB patient with his 4 relatives(cisAB type father and three O type relatives) and one B(A)02/O1 donor with his 3 relatives[ B(A) type father/brother and O type mother). For B(A)02/O1 donor, the results of genetic testing were consistent with the serological results, as the paternal genotype was the same as that of the proband, the younger brother was B(A)02/O2, and the maternal genotype was O1/O2. 【Conclusion】 The cisAB and B(A) blood groups are often indistinguishable by serological phenotypes and require genetic confirmation. CisAB pedigree investigation revealed 2 cases of cisAB blood type and B(A) pedigree investigation revealed 3 cases of B(A). The genotyping of cisAB and B(A) in this region were cisAB01/O2, B(A)02/O1, B(A)02/O2, B(A)04/O1 and B(A)04/O2. B(A)and cisAB subtypes can be accurately identified through genetic testing and pedigree investigation, which can provide a reliable basis for blood transfusion selection and ensure the safety of clinical blood transfusion.
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Objective To discuss the clinical characteristics of yon Hippel-Lindau (VHL)syndrome and the significance of genetic test for this disease.Methods Patients with VHL disease from 3 different families were reviewed from August 1985 to October 2017.The study was including clinical family survey and VHL-gene test on phylogenetic level.Totally 21 family members from 3 families were investigated,consisting of 14 males and 7 females with average age of 48.6 (5-70)years when analyzed.There were 8 patients with VHL disease comprising 5 males and 3 females with average onset age of 31.5 (9-67) years.Results The proband of pedigree one (VHL-Ⅱ C) was diagnosed as pheochromocytoma (PHEO) of right adrenal gland at 18 years old and underwent adrenalectomy,and her son was diagnosed with PHEO of bilateral adrenal glands with diagnostic age of 9 years old and received bilateral adrenalectomy sequentially.Her niece was diagnosed as PHEO of bilateral adrenal glands at 28 years old and received bilateral adrenal-sparing surgery simultaneously.Genetic analysis revealed a heterozygous mutation located at the third exon of the VHL gene (c.482G ＞ A).The proband of pedigree two (VHL-Ⅱ B) was diagnosed as right PHEO,bilateral multiple renal clear cell carcinoma (RCC),multiple pancreatic cysts and bilateral epididymal cystadenoma,and he received right adrenalectomy,right partial nephrectomy at 25 years old and delayed eystadenoma excision.His younger brother was also diagnosed as bilateral,pancreatic multiple cysts and bilateral epididymal nodules at 27 years old,and underwent right radical nephrectomy.Genetic analysis revealed a heterozygous mutation located at the first exon of the VHL gene (c.233A ＞ G).The proband of pedigree three (VHL-Ⅱ B) was diagnosed with central nerve system hemangioblastomas (CNS-HB) at 35 years old and received external beam radiotherapy.His elder sister was diagnosed as CNS-HB at 43 years old and received surgery.His father was diagnosed as right PHEO,bilateral RCC,bilateral multiple renal and pancreatic cysts and pancreatic neuroendocrine tumors at 67 years old.He received right adrenalectomy and partial nephrectomy.Genetic analysis showed a heterozygous mutation located at the third exon of the VHL gene(c.500G ＞ A).In addition,two cases (F2-Ⅲ 1 and F3-ⅣV1) were found to be asymptomatic VHL gene carriers by genetic screening.8 patients were followed up for an average of 9.8 (2-32) years.The symptoms were stable and no local recurrence or distant metastasis was found after operation.In this study,no CNS-HB was found in patients within family 1 and family 2,and RCC in 3 patients within family 2 and family 3 were low grade.Conclusions The clinical manifestations of VHL disease are diverse.RCC and CNS-HB are not present in all patients with the disease.PHEO is the only manifestation in patients with VHL-ⅡC.It is necessary to inform the members of VHL syndrome family for genetic test.Genetic test combined with clinical screening can facilitate differential diagnosis for VHL syndrome and other hereditary urological diseases.
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Background Leber hereditary optic neuropathy (LHON)is a mitochondrial DNA (mtDNA)hereditary disease,so it is significant to understand the influence of DNA mutation on the occurrence of LHON.Objective This survey was to evaluate the role of mtDNA mutation in the development of LHON.Methods This survey study was approved by the Ethic Committee of Wuhan General Hospital of Guangzhou Military Command and written informed consent was obtained from each subject before the relative medial examination.Seventy-two matrilineal relatives from a family with LHON were collected for a pedigree analysis and mutation screening.Regular eye examination was performed on 11 patients,13 mutant gene carriers and 49 individuals with normal phenotype,and the degree of visual damage was graded as follows: ＞0.3 was normal,0.1-0.3 was mild damage,＜0.05-0.1 was moderate damage,＜0.02-0.05 was severe damage and ＜0.01 was very severe damage.Clinical characteristics of LHON was evaluated.The periphery blood sample of 2-4 ml was collected from individuals to separate the mononuclear cells,and the mtDNA was extracted by modified high salt method.MtDNA was amplified by PCR and the mutation loci was sequenced.Results PCR amplification product sequencing of mutant gene showed that both G11778A and T14502C mutations were detected in 24 of 72 matrilineal relatives,but only 11 of 24 carriers developed LHON.No abnormal clinical findings were seen in the 13 carriers,showing a less 50％ penetrance in this family.There was no G11778A or/and T14502C mutation in the normal phenotype individuals of this family.The onset age for vision impairment in 11 affected matrilineal relatives varied from 8 to 50 years old,with the mean age of 24.36 years old,showing a significantly lower age than that of the 13 carriers (5-72 years old,mean 40.38 years old) (t =2.102,P=0.049).Conclusions This study suggests that the Gl1778A and T14502C mutation in mitochondrial DNA is one of causes in the development of LHON.The primary G11778A mutation together with T14502C mutation in mtDNA is a factor for the occurrence of LHON,hut it is not sufficient to the development of LHON.An effective “second hit” process will play an inducing role for LHON.
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One patient with congenital epidermolysis bullosa combined with pachyonychia was reported, and followed up for 11 years. The pedigree of the patient was investigated, and the loci of HLA-A, B antigens of 16 members of the pedigree were determined. The results showed that there were 60 members in the pedigree, among them 24 members had different kinds of skin diseases : 4 cases of epidermolysis bullosa combined with pachyonychia, 9 eases of albopapuloid form of epidermolysis hullosa combined with pachyonychia, 2 cases of icthyosis and 9 cases of pachyonychia simplex. The result of HLA-A, B antigens analysis of 16 members of the pedigree showed no evidence of the genes of these patients linked with HLA-A, B antigens.