摘要
目的探索建立一种简便、快速、较低成本的用于孕妇外周血胎儿有核红细胞(FNRBC)富集的实验方法。方法采集18名孕周在8～16周的孕妇外周血10ml,经Percoll不连续密度梯度离心初步分离后,运用Keihuaer抗酸染色法对玻片上的细胞进行染色标记,显微镜下观察、辨别FNRBC的特殊染色及形态,对阳性涂片者进一步行胎儿血红蛋白(HBF)特异性抗体标记以证实其胎源性。结果经Keihuaer抗酸染色处理,阳性FNRBC的胞浆染色呈深红色,核蓝色,而母亲的有核红细胞胞浆不着色。18名孕妇外周血中有14名可检出FNRBC,平均1～6个阳性FNRBC/病例,阳性检出率为77.8%。14例FNRBC阳性玻片经HBF抗体标记亦证实相同结果,而4例Keihuaer抗酸染色阴性病例亦表现HBF标记阴性,两者检出符合率100.0%。结论密度梯度离心结合Keihuaer抗酸染色法用于富集FNRBC具有较高的特异性,实验操作方法简便、快速,有望用于无创性产前基因诊断的推广应用。
Objective To investigate and establish a simple, rapid and lower-cost method to enrich fetal nucleated red blood celI（FNRBC） in peripheral blood of pregnant woman. Methods About 10 ml of peripheral blood sample from 18 pregnant women with gestational weeks arranged from 8 to 16 was collected, and then subjected to the discontinuous density gradient centrifugation. After the primary cell isolation, Keihuaer acid-fast staining method was used to mark the isolated cells previously spread on the slides. The special morphocytology and colour staining of FNRBC could be distinctly distinguished from the surrounding mother cells. Following that, the special antibody against fetal hemoglobin was further used to validate the fetus-originated characteristics for the positive FNRBC staining slides. Results After dealing with Keihuaer acid-fast staining, the cytoplasm of positive FNRBC was stained to be scarlet,nucleolus to be blue,while the cytoplasm of mother cells were colorless. The positive FNRBC were detected in 14 of 18 healthy pregnant women, average 1 to 6 cells per case,the positive rate was 77.8 %. The same experimental results was further validated with the special antibody against fetal hemoglobin immunocytochemical staining,the diagnose accordance rate was 100. 0%. Conclusions The density gradient centrifugation,together with Keihuaer acid-fast staining were highly special for the enrichment of FNRBC in peripheral blood of pregnant woman. The practical operation of this method was simple, fast and the future investigations would hopefully prove its worth in the clinical application of noninvasive prenatal diagnosis of gene disorders.
出处
《临床输血与检验》
CAS
2009年第3期207-210,共4页
Journal of Clinical Transfusion and Laboratory Medicine
基金
安徽省科技攻关项目(No.06013125B)资助
