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机构地区:[1]四川宜宾市第二人民医院,四川宜宾644000
出 处:《国际检验医学杂志》2014年第2期146-147,151,共3页International Journal of Laboratory Medicine
基 金:四川省卫生厅基金资助项目(20100583);四川省宜宾市科技局基金资助项目(2010SF011)
摘 要:目的探讨不同羊水细胞处理方法对ABO血型基因鉴定的影响。方法选取孕16~25周年龄21~38岁的孕妇53例,每例采集羊水标本2份,一份进行羊水细胞培养2~5d,收集培养的羊水细胞;另一份直接离心分离羊水细胞,然后提取羊水细胞DNA,采用序列特异引物引导的PCR反应(PCR-SSP)基因检测技术鉴定羊水细胞的ABO基因型,待婴儿出生后,抽取其外周血或脐带血进行微柱凝胶ABO血型检测。结果羊水细胞培养法有5例未检出,检出率为90.5%(48/53);直接法中有9例未检出,其中有3例结果无法识别,结果检出率为83.0%(44/53);24例新生儿的外周血或脐带血的微柱凝胶ABO血型检测结果显示有1例无结果,与基因型的结果比对完全一致。结论羊水细胞可直接进行PCR-SSP检测ABO血型,细胞培养法优于直接分析法。Objective To explore the effect of different treatment methods for amniotic fluid cells on ABO gene identification by polymerase chain reaction with sequence-specific primers (PCR-SSP). Methods Fifty three pregnant women with gcstational age from 16 to 25 weeks were selected. To extract double amniotic fluid specimens,one was for in vitro cell culture and the other was for separating aminotic cell using centrifuge. ABO genotypes were detected by PCR-SSP after DNA was extracted from the amniot- ic cells. After fetus gave birth,peripheral blood was collected from these newborns and was analyzed by microcolumn gel agglutina- tion assay. Results Among the 53 specimens from cell culture method,ABO genotype of 5 cases were not detected. The detection rate was 90.5 % (48/53). There were nine cases that were not detected in direct centrifuge method group and the detection rate was 83.0% (44/53). Among them,results of 3 cases could not identify. Among the twenty-four neonatal peripheral blood or cord blood cases detected by microcolumn gel agglutination assay, there was one case that not detected. The results were same with that of PCR-SSP. Conclusion ABO genotype can be analyzed in amniotie fluid cells by PCR SSP,and the cell culture method is advanced than direct centrifuge method.
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