Genetic analysis of an individual of Ay serologic phenotype / 南方医科大学学报

<p><b>OBJECTIVE</b>To study the cause of weak expression of Ay antigens on the surface of erythrocytes and understand its molecular characteristics.</p><p><b>METHODS</b>Genotypic analysis of an individual of Ay serologic phenotype was performed using polymerase chain reaction with sequence specific primers (PCR-SSP). Sequence analysis of the 7 exons and some of the introns was carried out with ABI Prism 3100 DNA sequencer, and the Ay gene sequence was compared with A102 reference sequence.</p><p><b>RESULTS</b>The genotype of the Ay individual was A102/101. Sequence analysis identified a nt467C>T mutation, nt1009A/G heterozygosis and a nt517G-/C- deletion in the intron 5.</p><p><b>CONCLUSION</b>The molecular genetic background of the Ay phenotype is polymorphic. Our findings are not sufficient to explain the cause of weak expression of Ay antigens on the surface of erythrocytes in this individual.</p>

Comparison between genotyping and serological phenotyping in RhCE blood group / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To genotype the RHCE gene of Hans, Xinjiang's Uigurs and Kazakstans in China, and to compare the results of RHCE genotyping with that of RhCc/Ee phenotyping.</p><p><b>METHODS</b>RHCE genes of 98 Hans with RhD positive and 230 Hans, 72 Uigurs and 18 Kazakstans with RhD/RHD negative were genotyped with PCR-sequence specific primer (SSP) technique.</p><p><b>RESULTS</b>The results of RHE/RHe genotyping from samples with RhD positive and negative were in accord with that of phenotyping. It would result in 4.44% error using C-->G polymorphism at nt48 of RHCE gene to genotype RHCE, and 4.05% failure of detection using the 109 bp insertion to detectRHCE gene in Chinese Hans. The results of RHE/RHe genotyping in unrelated 72 Uigurs and 18 Kazakstans with RhD phenotype were consistent with that of phenotyping, and false positive and false negative were not found in genotyping in Uigurs and Kazakstans tested.</p><p><b>CONCLUSION</b>The results of RHE/RHe and RHc genotyping were correct with PCR-SSP and accordant with that of phenotyping. Using the C48G polymorphism in exon 1 of RHCE to genotype RHC gene would result in false positive resulting from RHc mutation at this locus, and using the 109 bp insertion to genotype RHC gene would result in false negative because of the absence of the 109 bp. Therefore it is necessary to genotype RHC gene using more than two polymorphic loci.</p>

Determination of human RHD gene rhesus box and its significance / 中国实验血液学杂志

The aim was to determine RHD zygosity, further to investigate genetic structure of RHD gene, and to predict hemolytic disease of newborn (HDN). The upstream box, downstream box, and hybrid box of RHD gene were determined by PCR-SSP with 4 primers under the same conditions. The results showed that only hybrid box could be determined in RHD(-)/RHD(-) homozygosity. All the upstream box, downstream box, and hybrid box could be determined in RHD(+)/RHD(-) heterozygosity, while upstream box and downstream box except hybrid box could be determined in RHD(+)/RHD(+) homozygosity. Out of 50 cases of RhD(+), 5 cases (10%) were RHD(+)/RHD(-) heterozygosity, and the others (90%) were RHD(+)/RHD(+) homozygosity. 54 cases (55.1%), 36 cases (36.7%) and 8 cases (8.2%) were RHD(-)/RHD(-) homozygosity, RHD(+)/RHD(-) heterozygosity, and RHD(+)/RHD(+) homozygosity respectively in 98 unrelated cases of RhD(-) Chinese Hans. 2 cases of weak D were proved to be RHD(+)/RHD(-) heterozygosity. Out of 16 D(el) types, the upstream box, downstream box, and hybrid box could be determined in 10 cases (37.5%) and the upstream box and downstream box except hybrid box could be determined in 6 cases. Results detecting of RHD 10 exons in above samples proved the correctness of the method. It is concluded that the method is suitable for clinical application with its simplicity and veracity. There are many noneffective RHD genes (44.9%) in Chinese Hans with RhD(-) phenotype.

Study on blood ABO typing in patients with autoimmune hemolytic anemia / 中国实验血液学杂志

To explore effect of autoantibody on identification of ABO and RhD blood group, the blood samples of 38 patients with autoimmune hemolytic anemia (AIHA) were identified by routine typing and typing after chloroquine elution test as well as PCR. The results showed that out of 38 patients with AIHA, 11 cases (31.6%) of ABO blood group were difficulty typed, indirect antiglobulin test were positive, and contradiction between cells typing and sera typing were observed. 1 case of RhD(-) was mistyped as RhD(+) and anti-D was found in its serum. The blood group of these cases were typed correctly by chloroquine elution test. It is concluded that blood group identification of patients with AIHA can be interfered by autoantibody, and the correct typing for blood group of these patients may be determined by using combination of several methods to ensure safe transfusion.

Separation and cryopreservation of cord blood mononuclear cells / 中国实验血液学杂志

The influencing factors on cord blood storage after collection and mononuclear cell separation as well as cryopreservation were studied. The mononuclear cell are separated from blood after blood collection, then cryopreserved and washed after thawed. Results showed that the cord blood kept at 4 degrees C or room temperature less than 24 hours after blood collection, mononuclear cell separated by hydroxyethylstarch and 2 centrifugations, mononuclear cell cryopreserved with 50% DMSO and autoplasma from cord blood as protectives and washing the cells after thawing. In conclusion, the optimal project in this study can effectively preserve cord blood mononuclear cells.