HLA-A*11:452N differs from A*11:01:01:01 by a single nucleotide exchange in exon 1.

HLA-B*58:01:40 differs from HLA-B*58:01:01 by a single nucleotide change in exon 3, 507 C- > T (codon 145.3 CGC- > CGT).

HLA-A*26:206:02N differs from A*26:01:01:01 by a single nucleotide exchange in exon 3.

Several cases of the hemolytic disease of the fetus and newborn (HDFN) caused by immunoglobulin G (IgG) anti-M antibodies have been reported, in which almost all the HDFN-associated anti-M were warmly reacting. Here we report two cases of severe HDFN associated with cold-reacting IgG anti-M. In both cases, pregnancy was terminated, in weeks 33 and 23 respectively, due to a diagnosis of fetal growth retardation (FGR). To our knowledge, these are the most severe HDFN cases caused by cold-reacting IgG anti-M.

OBJECTIVE:To analyze the molecular polymorphisms of CD36 among 58 blood donors with CD36 deficiency and compare with CD36 positive controls.METHODS:A total of 58 donors with CD36 deficiency during a screening conducted in the laboratory from September 2019 to December 2020 were enrolled as the test group, including 39 males and 19 females, while 120 platelet donors with CD36 positive were randomly selected as the controls, including 76 males and 44 females. All of the subjects were Han nationality. The PCR-SBT method was used to detect coding region of CD36 gene, and molecular mutations were compared with those CD36 positive controls.RESULTS:Among the 58 donors with CD36 deficiency, mutations appears in 32 individuals. The detection rate for type I was 71.43% (5/7), and type II was 51.92% (27/52), while among the 120 controls, mutations appears in 12 donors (10%). In the CD36 antigen-deficient donors, 16 variations were found, in which 329-330 del AC with the highest frequency accounted for 20.69%, followed by 1228-1239 del ATTGTGCCTATT(15.52%) and 1156 C>T(10.34%). Two variations, 198-205 del GATCTTTG and 220 C>T, led to premature termination of translation; four mutations, 329-330 del AC, 560 ins T, 1011-1049 39bp dupl and 1343-1344 ins TCTT, caused translation frame shift; 1228-1239 del ATTGTGCCTATT led to deletion of four amino acids (Ile-Val-Pro-Ile) at sites 410-413 of the peptide chain. The 1140 T>A and 1275 G>A were synonymous mutations, and the other 7 mutations resulted in the substitution of single nucleotide. The platelet expression in the donors of CD36 positive with 329-330 del AC or 1228-1239 del ATTGTGCCTATT mutation (heterozygote) was lower than those CD36 positive individuals without mutations (homozygote).CONCLUSION:Multiple gene mutations in the CD36 coding region may cause CD36 deficiency, and the heterozygous individuals with mutations may lead to CD36 antigen reduction or deletion. Mutation is not detected in 44.83% of CD36 deficient individuals, there may be some other reasons for the CD36 antigen deficiency.

BACKGROUND:The null phenotype in P1PK blood group, known as "p," is extremely rare in the whole world. Individuals of p phenotype spontaneously form anti-PP1PK isoantibody. Here, we report a case of p phenotype with naturally occurring anti-PP1PK isoantibodies in a Chinese individual.STUDY DESIGN AND METHODS:Serology tests, containing alloantibodies screening and identification, were conducted to demonstrate the phenotype in P1PK blood group. The genotype of A4GALT gene was identified by haplotypes separation and sequencing.RESULTS:The serological assay demonstrated the p phenotype of the proband, presenting with 1:64 titer of anti-PP1PK . The sequencing data revealed a compound heterozygote consisting of A4GALT*P1.01 with c.343A>T and a novel allele based on A4GALT*01N.05 with an addition polymorphism c.100G>A. The sequence of the novel allele has been submitted to GenBank and the accession number OM912503 was assigned.CONCLUSION:Our study demonstrates a case of naturally occurring anti-PP1Pk in a Chinese individual with p phenotype, which is based on compound heterozygosity including one novel allele. As the proband is a young lady, monitoring the titer of anti-PP1PK and early initiation of medical intervention are essential after her pregnancy.

目的 研究1例RhD血型鉴定部分凝集结果个体及其家系血清学表现和RHD基因.方法 通过血型微柱凝胶卡检测先证者ABO及RhD血型;盐水试管法检测先证者及其父母RhCcEe抗原;间接抗人球蛋白试验(indirect anti-human globulin test,IAT)及流式细胞术检测先证者RhD抗原.PCR序列特异性引物(PCR sequence specific primer,PCR-SSP)检测RHD基因以及RhD杂合型分析,基因测序方法分析RHD基因编码区序列.结果 血清学检测发现先证者血型为A型RhCcee,血型微柱凝胶卡、盐水试管法以及IAT法检测RhD抗原,结果呈部分凝集现象.流式细胞术结果显示先证者RhD抗原性减弱.经RHD基因编码序列分析发现,RHD基因第9外显子上的第1212位碱基发生C>A纯合突变,为RHD*weak D type 72的特征性突变点.家系调查显示,先证者父亲为O型RhCCDee,母亲为A型RhCcDee.父亲携带RHD*weak D type 72等位基因,基因型为RHD*weak D type 72/RHD+;母亲一条染色体缺失了全部的RHD基因,基因型为RHD+/RHD-.证明先证者分别从父亲和母亲遗传RHD*weak D type 72和RHD-等位基因,基因型为RHD*weak D type 72/RHD-.结论 发现了1例RHD*weak D type 72/RHD-基因型个体,丰富了RHD*weak D type 72变异型的研究数据.根据家系调查证明,RHD*weak D type 72等位基因由遗传获得,而非由个体基因变异形成.

Background:The HLA-E gene is a member of the HLA-I gene family. Its genetic polymorphism is regarded as associated with numerous diseases. Establishing a rapid and accurate detection method of disease-related SNP sites in HLA-E is particularly important.Methods:Blood samples from 226 healthy blood donors and 228 leukemia patients were collected, and DNA was extracted. Three typing methods based on PCR-sequence-based typing, TaqMan genotyping, and high-resolution melting curve were established to identify rs76971248 (G>T). The Chi-square test was used for statistical analysis by SPSS.Results:Three methods based on PCR-SBT, TaqMan genotyping, and HRM were all able to identify rs76971248. The software for analyzing the results of HLA-E sequencing was easy to use, and the results were accurate. The frequency of rs76971248 in different types of leukemia patients was significantly lower than that in healthy blood donors (p < 0.05). And the frequency of the G/G genotype in leukemia patients was significantly higher than that in healthy blood donors (p < 0.05).Conclusions:For the screening of known SNP sites in large-scale populations, among the three methods, the TaqMan genotyping method had the advantage of shortest time consumption, simplest operation, and greatest specificity, which was the most appropriate method for this experiment. The analysis software for HLA-E gene sequencing needed to be further optimized. rs76971248 had a protective effect against leukemia. And the G/G genotype was a risk factor for leukemia.

The authors have disclosed no conflicts of interest. Appendix S1. Supporting information. Please note: The publisher is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.

目的 研究深圳地区血清学弱D表型献血者RhCcEe表型与RHD基因型特点.方法 收集38例献血者血液样本,抗-D抗体(IgM/IgG)检测RhD抗原.采用IgM抗-C、抗-c、抗-E、抗-e抗体检测RhCcEe抗原.利用PCR-SSP方法分析RHD基因10个外显子,必要时对RHD基因的所有外显子进行直接测序.统计学方法分析RhCcEe表型与RHD基因型之间的相关性.结果 38例血清学弱D表型献血者中发现8种已报道的等位基因,RHD*weak partial 15(15/38)、RHD*DEL1(11/38)、RHD*DVI.3(5/38)、RHD*weak D type 61(1/38)、RHD*weak D type 95(1/38)、RHD*DCC(1/38)、RHD*DFR1(1/38)、RHD*weak D type 50(1/38)、RHD*weak partial 15/RHD*DEL1杂合(1/38).其中RHD*weak D type 50在中国汉族人群中首次报道.另外,发现了1例新的RHD等位基因RHD*IVS9-1C(c.1228-1G>C).该等位基因的序列数据已提交GenBank,登记号为MT755965.经相关性分析,RHD*weak partial 15与RhE抗原相关系数为0.727(P<0.01),RHD*DEL1与RhC抗原相关系数为0.645(P<0.01).结论 深圳地区血清学弱D表型献血者的RHD基因结构呈现多态性,主要为RHD*weak partial 15、RHD*DEL1、RHD*DVI.3,并存在其他稀有的基因型;RHD*weak partial 15与R h E抗原、RHD*DEL1与RhC抗原存在显著正相关.

The polymorphism of MNS blood group antigens is determined by the amino acid sequences of glycophorin A (GPA) and glycophorin B (GPB). Genetic mutations in their coding genes GYPA and GYPB might lead to the changes of amino acid and influence the expression of antigens. The determinants of the polymorphism of M and/or N antigens are the 1st and 5th amino acids of mature GPA protein (residues 20 and 24 counting from the translation-initiating site), while the polymorphism of S and/or s antigens are determined by the 29th amino acids of mature GPB. 2 The allelic frequency of GYPA and GYPB demonstrates regional differences in human genetics 3 We are committed to exploring the genetic polymorphism of these genes. Here, we describe a novel mutation, c.125G > C, on the GYPA*01 allele in a 28-year-old Chinese male, which caused absence of M antigen in red blood cells (RBCs).

The authors have disclosed no conflicts of interest. Fig. S1 Genotype. The top chromatogram (direct sequencing) shows the heterozygous sequence (T/C at nt.917). The lower two chromatograms (after cloning and sequencing) show separate nucleotide sequences from each allele. Please note: The publisher is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.

目的 总结并探讨深圳地区最近10年临床疑难交叉配血(标本)不合原因,为提高输血疗效提供解决方案.方法 收集2011年～2020年深圳地区医疗机构送本中心做疑难血型鉴定的1770例临床标本,分析其交叉配血不合的原因,确定患者血浆中意外抗体的类型,通过分类变量的统计学方法对抗体做总体率评估.以同型输血结合相容性相输的原则来提高输血的安全性.结果 1)1770例疑难配血标本来源于956名患者,其中2011年～2015年疑难配血患者人均配血次数为1.32(307/232);自2016年始,疑难配血患者人均配血次数由2016年的1.27(103/81)上升至2018年的2.23(286/128),并在之后2年保持稳定.2)在956名其交叉配血不合患者中,由自身抗体或/和同种抗体阳性的比例为90.38％ (864/956),包括42.26％ (404/956)自身抗体合并同种抗体阳性、20.71％(198/956)单纯自身抗体阳性、27.41％(262/956)单纯同种抗体阳性;检出特异性同种抗体20种,分属于8个血型系统,其中Rh血型的55种抗体的频率达到70.82％(551/778)[抗-E(37.15％)＞抗-c(20.95％)抗-C(5.27％)=抗-e(5.27％)＞抗-D(2.19％);其后依次为MNS[11.40％(112/778)]、Kidd[5.66％(44/778)]、Leiws[3.21％ (25/778)]、Duffy[1.80％(14/778)]、Diego[1.03％ (8/778)]、P1[0.39％ (3/778)]、H[0.26％ (2/778)]血型系统的同种抗体.3)年龄＜20岁的多次输血患者中有86％ (37/43)为地中海贫血患者,均检测到自身抗体或红细胞同种抗体1～4种.结论 临床交叉配血不合的原因主要是由输血或妊娠免疫导致的同种抗体、自身免疫性疾病如自身免疫性溶血性贫血引发的自身抗体干扰配血所致;临床医疗机构如遇交叉配血不合,应立即送至具有检测能力的血型参比实验室鉴定和配血,这对于保障临床安全用血与提高疗效至关重要.

Figure S1. Genotype. The top chromatogram (direct sequencing) shows the heterozygous sequence (delA at nt.36–39) of the proband. The lower two chromatograms (after cloning and sequencing) show separate nucleotide sequences from each allele. Please note: The publisher is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.

Figure S1. Sequencing results of exon 10 and flanking intron region of RHD. (A) Sequence analysis result of the proband. The arrow points to RHD c.1228-1G>C. (B) Sequence analysis result of the daughter. The arrow points to RHD c.1228-1G/C. Figure S2. Family pedigrees of the proband with a RHD c.1228-1G>C allele. The D haplotype carrying a RHD c.1228-1G>C allele was found in the proband (black background) and also detected in the daughter. Please note: The publisher is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.