Genetic Polymorphisms Of DEL Type In Chinese Han, Uyghur And Tibetan Population

Objective:1. To study comparatively on distribution characteristics of RhD-negative phenotypes in Chinese Han, Uyghur and Tibetan population.2. To study on distribution characteristics of Rh phenotypes in DEL type in Chinese Han and Uyghur population and reaserch the relation of DEL type with RhCc.3. To research the gentic structure and characteristics of DEL type in Chinese Han and Uyghur population, to explore the relationship between the genetic polymorphism of DEL type and the expression quantity of D antigen.4. To explore common mutations types and gene combinations of DEL in Chinese Han and Uyghur, and to analyze the correlation between DEL and RhC antigen, as well as to establish the molecular detection method of DEL type for common Chinese population.5. To provide genetic polymorphism information of Chinese DEL type for international genetic database.Methods:1. Rh phenotypes were detected with monoclonal Anti-D, Anti-C, Anti-c, Anti-E and Anti-e antibodies. RhD-negative samples were screened with modified indirect antiglobulin test (IAT); The IAT-negative samples were detected with microcolum gel test of adsorption and elution assay.2. RHD1227A polymorphisms were detected by PCR-SSP method using specific primers, and results were verified with sequencing.3. RHD exon9was detected by PCR method using specific primers, and results were verified with sequencing.4. Upstream box, downstream box and hybrid box of RHD gene were detected by PCR-SSP method, and partial samples were verified with sequencing.5. Studty data were analyzed with SPSS18.0statistical software.Results:1.6kinds of Rh phenotypes were detected in222RhD-negative samples from random unrelated Chinese Han, which were CCee (4.95%)、Ccee (31.08%)、ccEE (0.90%)、ccEe (4.06%)、ccee (56.76%) and CcEe(2.25%), respectively;3kinds of Rh phenotypes were detected in55RhD-negative samples from random unrelated Uyghur in Xinjiang, which were CCee (5.45%)、Ccee (3.64%) and ccee (90.91%), respectively. The distribution of Rh phenotypes between Han, Uyghur and Tibetan RhD-negative population existed statistically different (x2=17.04P=0.000), and the same difference between Han and Uyghur population, Uyghur and Tibetan population (χ2=25.031, P=0.000; χ2=21.761, P’=0.000) were also seen. However, there was not different between Han and Tibetan population(χ2=1.337, P’=0.720); The distribution of the Cc phenotypic between Han and Uyghur RhD-negative individuals existed statistically different too (χ2=19.757, P=0.000).2.222RhD-negative samples from random unrelated Chinese Han were phenotyped as74Cc,11CC and137cc phenotypes, respectively. Of which43cases of DEL type were detected, and their distributions of Cc were36(83.72%),4(9.30%),3(6.98%), respectively.55RhD-negative samples from random unrelated Uyghur persons were phenotyped as2Cc,3CC and50cc phenotypes, respectively, of which only one DEL type was detected; DEL type was not detected in7Tibetan RhD-negative samples.3. Out of222RhD-negative samples of Chinese Han,157samples (70.72%) were detected carrying hybrid box of RHD gene, which meant these samples had RHD/RHD genotypes,60samples (27.03%) were detected to have upstream box, downstream box and hybrid box, and only5samples (2.25%) were detected to have upstream box and downstream box; Out of55RhD-negative samples of Uyghur individuals in Xinjiang,54samples (98.18%) were detected to have hybrid box, and one sample has upstream box and downstream box;6(85.71%) out of7RhD-negative samples of Tibetan were detected to have hybrid box, the rest one was detected to have upstream, downstream and hybrid box. The genotype distribution of RhD-negative samples among three nationas existed statistically different (F=27.389, P=0.000). There existed statistically difference of RHD genotype distribution between Han and Uyghur population with RhD-negative (F=26.808, P=0.000), while it showed no significant difference between Chinese Han and Tibetan population (F=0.816, P=0.242); The frequencies of RHD/RHD genotype existed statistically different between Han and Uyghur population (χ2=18.315, P=0.000).4. Upstream box, downstream box and hybrid box of RHD gene could be detected in all10weak D samples from Chinese Hans. Out of43samples from Chiese Hans with DEL type,41could be detected upstream box, downstream box and hybrid box of RHD gene, while one sample could be detected upstream box and downstream box. One Uyghur sample could be detected upstream box and downstream box. RHD1227G>A polymorphisms were detected in43cases of DEL type from random unrelated Chinese Hans by PCR-SSP, of which one sample was RHD1227G,3samples were1227G/A and39samples were1227A, respectively. Combining with Rhesus box test results, their gene combinations were RHD1227G/-, RHD1227G/A, RHD1227A/-or RHD1227A/A genotype, respectively; The unique Uyghur DEL sample was1227A/A genotype. RHD1127A allele could not be detected in all homozygous RHD/RHD samples.40cases (93.02%) of DEL type samples with RHD1227A or RHD1227A/G genotype carried RhC antigen.Conclusions:1. The distribution of Rh phenotype in Chinese Han、Uyghur and Tibetan population with RhD-negative existed statistically different. The main phenotype was ccee, and which was significantly higher in Uyghur population than that in Chinese Han population.2. In Chinese Han, Uyghur and Tibetan population, RHD genotypes with RhD-negative phenotypes were mainly RHD-/RHD-homozygotes. RHD genotypes in most of the Chinese Hans with DEL types were mainly RHD+/RHD-heterozygote, which meant that all DEL type individuals had RHD gene.3. RHD1227A allele was common in Chinese Han and Uyghur with DEL type, which showed the relationship with RhC antigen.4. The detection method of common Chinese DEL type was established, and it is helpful to research on genetic characteristics of DEL type in China.