[Study on molecular genetic structure of Ael blood subgroup].

Objective: To study the ABO allele molecular characteristics of Ael blood subgroup.

Methods: Five individuals of diagnosed as Ael blood subgroup were subjected to PCR amplify ABO alleles using four pairs of sequence-specific primers. Exon 6 and exon 7 at ABO locus of all samples were sequenced.

Molecular genetic analysis for Ax phenotype of the ABO blood group system in Chinese.

Background And Objectives: To elucidate the molecular genetic background of the Ax phenotype in the Chinese population.

Materials And Methods: The ABO genes of eight Ax phenotype samples, four Ax and four AxB, were amplified by polymerase chain reaction (PCR) and were cloned, along with those of 10 random A(1) Chinese subjects. We analysed the ABO gene transcript structure and the sequences of two exons and one intron at the ABO locus.

[Generation of RHD-CE(2-9)-D allele by gene conversion in cis].

Previously a few Rh-negative individuals in Caucasian and Chinese were found existing exons 1 and 10 of RHD through genomic DNA testing. The molecular mechanisms, however, remain disputed. In this study, 2 individuals carrying RHD positive, D antigen negative allele (with exon 1 and 10 of RHD) and their mRNA were investigated by using reverse transcriptase PCR (RT-PCR) and sequencing through one pair of specific primers for 5'- and 3'-non-coding region of RHD, taking a Rh-positive (Ccee) sample as control.

[Identification of a novel B variant allele in a Chinese Han individual with B subgroup].

Objective: To study the molecular genetic background of B subtype in Chinese Han population and identify novel allele at the ABO locus.

Methods: Ten samples from randomly selected blood donors of normal B phenotype used as control, and six samples from individuals diagnosed as B subgroup by serological tests were genotyped by sequence specific primer polymerase chain reaction and direct DNA sequencing at the exons 6 and 7 of ABO gene. The exons 6 and 7 and the intervening intron 6 of the B allele from each B subgroup sample were analyzed by cloning and haplotype sequencing.

[Molecular genetic analysis for the A3 alleles].

To study four A(3) subgroup samples identified by serologic tests, among which two belong to a family, three were A(3) subgroup, one was A(3)B subgroup. All four samples were genotyped by PCR-SSP method, and the nucleotide sequences of Exon 6, Exon 7 and part introns at the ABO locus for these samples were detected by ABI Prism 3100 DNA sequencer. Comparison with the consensus of A101 was performed.

[Study on the simultaneous genotyping of human platelet antigens of 1, 2, 3, 4, 5, 6 system by PCR-SSP and its applications].

To set up the simultaneous genotyping of human platelet antigens of 1,2,3,4,5,6 system by PCR-SSP assay and use the genotyping method for the study of platelet antigens. In this study, 18 sequence-specific primers were designed and synthesized. The annealing temperature for all sequence-specific primer pair, the concentration of each primer pair and the concentration of Mg2+ were adjusted to the optimum so that HPA-1 to 6 systems could be amplified simultaneously under the same PCR cycling parameters.

[Genetic polymorphisms of 6 Y-chromosome specific STR loci in the southern Chinese Han population and its application in forensic science].

To study the genetic polymorphisms of six Y-chromosome specific STR loci in the southern Chinese Han population and apply it in forensic science, six Y-STR loci were amplified by multiple PCR and the PCR products were detected by using ABI Prism 377 Sequencer. The haplotype frequencies at 6 Y-STR loci were determined in a total of 204 unrelated males from southern Han population of China. Ninety-three father/son pairs with demonstrated paternity and thirty-eight non-paternity father/son pairs were detected by using our Y-STR system.

[Study of HLA polymorphism in the 6965 Han bone marrow registry donors].

Objective: To analyze human leukocyte antigen (HLA) polymorphism and search for new alleles in Chinese Han population bone marrow registry donors.

Methods: DNA-based HLA genotyping methods were used including PCR-SSP, BST and molecular cloning.

Results: A total of 6965 unrelated donors, 4707 from South China origin and 2258 from north, were typed for HLA-A, B, and DRB1 loci.

[A new PCR method for direct determination of RHD zygosity].

Objective: To establish a direct method for the determination of RHD zygosity.

Methods: Two pairs of primers were designed specific for hybrid Rhesus box and exon 1 of RHD, respectively. Combined with a pair of internal control primers, a new dual-tube PCR method was established.

Whole exon 5 and intron 5 replaced by RHCE in DVa(Hus).

The DVa(Hus) was previously investigated through cDNA analysis, which revealed an RHD-CE(5)-D hybrid allele. However, the 5' and 3' breakpoints remain unknown. In this article, gene recombinations between the RHD and RHCE alleles were investigated by a combination approach of a sequence-specific primer PCR (PCR-SSP) and an RHD full-length coding region sequencing method on two Chinese subjects with weak D phenotypes.

[Applications of RHD zygosity test through polymerase chain reaction for prediction of fetus Rh D-positive phenotype].

Objective: To explore a molecular method for the determination of RHD zygosity.

Methods: Two pairs of primers were designed specific for downstream rhesus box and hybrid rhesus box according to the sequences in GenBank. Together with a pair of internal control primer, a dual-tube polymerase chain reaction (PCR) method was established for determination of the RHD zygosity.

Identification of a novel allele HLA-B*5610 in a Chinese potential bone marrow donor.

A novel HLA-B allele, B*5610, has been identified in a potential bone marrow donor, his mother and brother using DNA-based typing and molecular cloning methods. The B*5610 allele differs from the closest matching HLA sequence of B*5602 by two nucleotide substitutions in exon 3, 559 C-->A and 560 T-->C, resulting in an amino acid change from Leu (CTG) to Thr (ACG) at codon 187. This new allele was segregated together with A*24020101 and DRB1*140101 in the proband's family.

Identification of a new HLA allele, A*1114, in a Chinese family.

A novel HLA-A allele, A*1114, was initially detected in two generations of a Chinese family by unusual polymerase chain reaction based sequence-specific primers ( PCR-SSP) reaction patterns and ambiguous sequence-based typing (SBT). Molecular cloning and sequencing analysis indicated that this new allele differs from HLA-A*1102 by three nucleotide substitutions in exon 3, 524 A-->G, 526 G-->C, and 527 C-->G, thus changing codon 175 from His to Arg (CAT-->CGT) and codon 176 from Ala to Arg (GCG-->CGG). Segregation analysis showed that the proband inherited his mother's HLA haplotype A*1114, B*5801, DRB1*1405.

Cloning and complete sequence of a novel HLA-A null allele, A*0253 N, with a termination codon generated by a C to G mutation in exon 2.

A novel HLA-A null allele, A*0253 N, has been identified in two generations of a Chinese family using combined serological and molecular cloning approaches. Full-length genomic DNA sequencing indicated that this new allele differs from HLA-A*02011 by a single C to G substitution at nucleotide position 324 in exon 2. This mutation results in an amino acid change from a tyrosine codon to a stop codon at position 108.