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一种检测母血中胎儿RNA的半定量微阵列方法。

A semi-quantitative microarray method to detect fetal RNAs in maternal plasma.

作者信息

Ge Qinyu, Liu Quanjun, Bai Yunfei, Wen Tian, Lu Zuhong

机构信息

State Key Laboratory for Bioelectronics, Southeast University, Nanjing, China.

出版信息

Prenat Diagn. 2005 Oct;25(10):912-8. doi: 10.1002/pd.1253.

Abstract

OBJECTIVES

To set up a semi-quantitative microarray method for the detection of fetal RNAs in maternal plasma.

METHODS

We developed a semi-quantitative microarray method for the detection of placental RNA in maternal plasma. Firstly, the selected fetal RNAs were linearly amplified from the maternal plasma and then fluorescently labeled as the target DNAs. Finally, the targets were hybridized and detected by capturing DNA probes on a microarray slide. Two genes of beta subunit of human chorionic gonadotrophin (beta-hCG) and zinc finger gene on the Y chromosome (ZFY) were assayed with the microarray, and beta actin gene was used as an internal standard. Eighty-five pregnant women in the first trimester and the third trimester of gestation joined in this experiment, 14 of them also sampling in 36 h after delivery for the same assay. Real-time quantitative PCR was performed for comparison.

RESULTS

It was found that the mRNA level of beta-hCG decreased with the increasing of gestation age, and it was much higher in the carriers of the female fetus than in the carriers of the male fetus in the first trimester of gestation, which was consistent with the real-time quantitative PCR results. The results also reveal that delivery would result in the clearance of fetal mRNA in maternal plasma.

CONCLUSIONS

The results suggest that the semi-quantitative microarray method has great potential as a high-throughput assay in prenatal diagnosis and clinical laboratory.

摘要

目的

建立一种用于检测孕妇血浆中胎儿RNA的半定量微阵列方法。

方法

我们开发了一种用于检测孕妇血浆中胎盘RNA的半定量微阵列方法。首先,从孕妇血浆中线性扩增所选胎儿RNA,然后将其荧光标记为靶DNA。最后,通过在微阵列载玻片上捕获DNA探针来杂交和检测靶标。用微阵列检测人绒毛膜促性腺激素β亚基(β-hCG)和Y染色体上的锌指基因(ZFY)这两个基因,并以β肌动蛋白基因作为内标。85名孕早期和孕晚期的孕妇参与了本实验,其中14人在分娩后36小时也进行了相同检测的采样。进行实时定量PCR以作比较。

结果

发现β-hCG的mRNA水平随孕周增加而降低,且在孕早期女性胎儿携带者中比男性胎儿携带者中的水平高得多,这与实时定量PCR结果一致。结果还显示分娩会导致孕妇血浆中胎儿mRNA的清除。

结论

结果表明,半定量微阵列方法作为一种高通量检测方法在产前诊断和临床实验室中具有巨大潜力。

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