摘要
目的比较中国汉族和维吾尔族(维族)人群D_(el)型基因结构和遗传特点,了解二者D_(el)型常见突变类型及其基因组合情况。方法采用微柱凝胶法检测随机非血缘关系的中国222名汉族人和55名维族人的Rh D血型,Rh D(-)者用改良间接抗球蛋白试验(IAT)确认;单克隆抗体鉴定Rh表型;吸收放散微柱凝胶法检测D_(el)型;PCRSSP方法检测RHD上游、下游和杂合Rhesus盒;PCR扩增RHD第9外显子,测序验证;PCR-SSP检测RHD基因1227位点,测序验证。结果 222例随机非血缘关系汉族血清学Rh D(-)标本和55例随机非血缘关系维族血清学Rh D(-)标本分别检出43例和1例D_(el)型;汉族Rh D(-)标本中,RHD基因型为RHD-/RHD-和RHD+/RHD-、RHD+/RHD+者的比例分别为70.72%(157/222)和27.03%(60/222)、2.25%(5/222);维族Rh D(-)标本中,只有1例RHD基因型为RHD+/RHD+,其余98.18%(1/54)标本为RHD-/RHD-;汉族和维族Rh D(-)个体RHD基因型总体分布存在差异(P<0.05);汉族和维族Rh D(-)个体的RHD-/RHD-基因型呈明显差异(P<0.05);43例随机非血缘关系的汉族D_(el)型标本RHD基因1227位点检测:1例1227G、3例1227G/A和39例1227A,结合Rhesus box检测结果,基因组合情况分别为RHD1227G/-、RHD1227G/A、RHD1227A/-和RHD1227A/A。1例维族D_(el)型标本等位基因组合为RHD1227A/A。所有RHD-/RHD-纯合子样本均未检测到RHD基因1227位点。吸收放散试验为D_(el)型者93.02%(40/43)、等位基因为RHD1227A或A/G个体均携带C抗原。结论中国汉族和维族D_(el)型人群RHD基因型绝大部分是RHD+/RHD-杂合子,都存在RHD;中国汉族和维族D_(el)型等位基因以RHD1227A为主,且与C抗原相关联。
Objectives To research the genetic structure and characteristics of Deltype in China Han and Uyghur population,and to study frequent mutations types and gene combinations of Deltype. Methods Rh phenotypes of 222 Rh D-negative samples from random unrelated Chinese Han and 55 Rh D-negative samples of Uyghur individuals were detected with monoclonal Ig M antibodies. Deltypes were distinguished from the IAT-negative samples by microcolom gel test together with adsorption and elution assay. Upstream box,downstream box and hybrid box of RHD gene were detected by PCR-SSP method. RHD exon 9 was detected by PCR method using specific primers,and results were verified with sequencing. RHD1227 A polymorphisms were detected by PCR-SSP,and their results were also verified with sequencing. Results 43 cases of Del type were detected 222 Rh D-negative samples from random unrelated Chinese Han and 1 Deltype was found from 55 Rh Dnegative samples of Uyghur individuals. Among the Rh D-negative samples of Chinese Han,the proportion of genotypes of RHD- / RHD- and RHD ＋ / RHD-,RHD ＋ / RHD ＋ respectively 70. 72%（ 157 /222） and 27. 03%（ 60 /222）,2. 25%（ 5 /222）. Out of the Rh D-negative samples of Uyghur individuals,only one sample＇ s genotype was RHD ＋ / RHD ＋,the other98. 18%（ 1 /54） were RHD- / RHD-. The genotype distribution of Rh D-negative samples between Han and Uyghur population showed significant differences statistically（ P〈0. 05）. The frequencies of RHD- / RHD- genotype existed statistically different between Han and Uyghur population（ P〈0. 05）. RHD 1227 G〉 A polymorphisms were detected in 43 cases of Del type from random unrelated Chinese Hans by PCR-SSP,of which,one sample was RHD1227 G,3 samples were 1227 G / A and39 samples were 1227 A,respectively. Combining with Rhesus box test results,their gene combinations were RHD1227 G /-,RHD1227 G / A,RHD1227 A /- or RHD1227 A / A genotype,respectively; The unique Uyghur Delsample was 1227 A / A genotype. RHD1227 A allele could not be dete
出处
《中国输血杂志》
CAS
北大核心
2015年第12期1451-1454,共4页
Chinese Journal of Blood Transfusion
基金
国家自然科学基金(309715807)
